Peptidomimetic agents, synthesis and uses thereof

ABSTRACT

Compounds for use in synthesis of peptidomimetic agents; synthesis of peptidomimetic agents; peptidomimetic diagnostic and therapeutic agents; and uses of the compounds and peptidomimetic agents in drug discovery, diagnosis, prevention and treatment of diseases are described.

This application claims the benefit of U.S. Provisional Application No.62/845,611, filed on May 9, 2019, hereby incorporated by reference.

FIELD OF THE INVENTION

The present invention is directed to compounds or synthons for use insynthesis of peptidomimetic agents; peptidomimetic agents; synthesis ofpeptidomimetic agents; and uses of these synthons and peptidomimeticagents in drug discovery, diagnosis, prevention and treatment ofdiseases.

BACKGROUND OF THE INVENTION

The in vitro and in vivo stability, in vitro and in vivo half-lives andefficacy of peptides are limited, e.g., by the rate of hydrolysis andenzymatic degradation of the peptides. As the result of the rapidhydrolysis and enzymatic degradation, a number of peptides have ahalf-life that is too short to be used in drug discovery, diagnosis,prevention and treatment of diseases. For example, the in vivo half-lifeof bradykinin is less than 1 minute, and the the in vivo half-life oftetramer peptide FSSE is less than 5 minutes.

In addition, as the result of the rapid hydrolysis and enzymaticdegradation, a number of peptides are not stable enough to be used indrug discovery, diagnosis, prevention and treatment of diseases. Forexample, the amount of endomorphin-2 (e.g., AUC (O.D.254)) in mouseserum at 2 hours after endomorphin-2 was placed in mouse serum is about95% less than the amount of endomorphin-2 originally placed in the mouseserum. Similarly, the amount of bradykinin (e.g., AUC (O.D.220)) inmouse serum at 3 hours after bradykinin was placed in mouse serum isabout 97% less than the original amount bradykinin placed in the mouseserum.

Moreover, as the result of the rapid hydrolysis and enzymaticdegradation, a number of peptides are not orally bioavailable or have anoral bioavailability that is insufficient for formulation into an oraldosage form.

There is a need for analogues of peptides that are more stable tohydrolysis and enzymatic degradation and/or are more active than theoriginal peptides.

There is also a need for building blocks or synthons for the synthesisof analogues of peptides and methods of synthesizing analogues ofpeptides that are more stable to hydrolysis and enzymatic degradationand/or more active than the original peptides.

SUMMARY OF THE INVENTION

The present invention provides building blocks or synthons for synthesisof analogues of peptides (“peptidomimetic agents”). Peptidomimeticagents assembled from these building blocks or synthons are moreresistant to hydrolysis and enzymatic degradation than the originalpeptides. As compared to the original peptides, the peptidomimeticagents contain one or more nitrogen(s) instead of one or moreα-carbon(s) of the original peptides, e.g., at the N-termini of thepeptide (i.e., the first residue of the peptide), at the second residueof the peptide, the C-termini of the peptide (i.e., the last residue ofthe peptide), the residue covalently bound to the C-termini of thepeptide, and/or at another residue of the peptide (e.g., at the site ofhydrolysis of the peptide). The replacement of one or more α-carbon(s)with one or more nitrogen(s) results, e.g., in an improved stability ofthe peptidomimetic agent to hydrolysis and enzymatic degradation, ascompared to the original peptide. The use of the building blocksdisclosed herein may allow, e.g., for synthesis of peptidomimetic agentsthat have longer in vitro and in vivo half-lifes than the originalpeptides, and correspondingly have utility in treating humans for a widevariety of diseases and conditions; and/or for synthesis ofpeptidomimetic agents at a reduced cost and/or increased yields, ascompared to conventional syntheses which do not use these buildingblocks. The synthesis of peptidomimetic agents using the building blocksdisclosed herein, the peptidomimetic agents and uses of thepeptidomimetic agents in drug discovery, diagnosis, prevention andtreatment of diseases are within the scope of the present invention.

It is an object of the invention to provide compounds or synthons foruse in synthesis of peptidomimetic agents.

It is an additional object of the invention to provide peptidomimeticagents for use in drug discovery, diagnosis, prevention and treatment ofdiseases.

It is yet an additional object of the invention to providepeptidomimetic diagnostic and therapeutic agents.

It is a further object of the invention to diagnose, prevent and treatdiseases with peptidomimetic agents.

In connection with the above objects and others, the invention isdirected in part to the compounds of Formula (IA):

wherein A is N-phthalimidyl or NR₁R₂,

R₁ is H,

R₂ is tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, or

R and R₁ are connected and together form a side chain radical ofproline;

X is imidazolyl or benzotriazolyl; and

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, threonine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, serine, and glutamine. Imidazolyl andbenzotriazolyl may be unsubstituted or substituted with one or more ofthe following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl(e.g., —CF₃, —CHF₂, —CH₂F, —CBr₃, —CHBr₂, —CH₂Br, —CCl₃, —CHCl₂,—CH₂Cl), —NH₂, or NH₃. In certain embodiments, the imidazolyl andbenzotriazolyl are substituted with —CF₃. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In certain embodiments, R is selected from thegroup consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, threonine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, asparagine, andglutamine. Compounds of Formula (IA) could be used in drug discovery,diagnosis, prevention and treatment of diseases, or as building blocksfor synthesis of peptidomimetic agents, e.g., for use in drug discovery,diagnosis, prevention and treatment of diseases.

In certain embodiments, a compound of Formula (IA) is a compound inwhich

A is N-phthalimidyl or NR₁R₂,

R₁ is H,

R₂ is tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl;

X is imidazolyl or benzotriazolyl; and

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, threonine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, and glutamine. Imidazolyl andbenzotriazolyl may be unsubstituted or substituted with one or more ofthe following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl(e.g., —CF₃, —CHF₂, —CH₂F, —CBr₃, —CHBr₂, —CH₂Br, —CCl₃, —CHCl₂,—CH₂Cl), —NH₂, or NH₃. In certain embodiments, the imidazolyl andbenzotriazolyl are substituted with —CF₃. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In certain embodiments, R is selected from thegroup consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, threonine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, asparagine, andglutamine.

In certain embodiments, a compound of Formula (IA) is a compound inwhich

A is N-phthalimidyl or NR₁R₂,

R₁ is H,

R₂ is tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl;

X is imidazolyl substituted with —CF₃, —CHF₂, —CH₂F, —CBr₃, —CHBr₂,—CH₂Br, —CCl₃, —CHCl₂, —CH₂Cl; and

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, and glutamine. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In certain embodiments, R is selected from thegroup consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, threonine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, asparagine, andglutamine.

In certain embodiments, a compound of Formula (IA) is a compound inwhich

A is N-phthalimidyl or NR₁R₂,

R₁ is H,

R₂ is tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl;

X is benzotriazolyl substituted with —CF₃, —CHF₂, —CH₂F, —CBr₃, —CHBr₂,—CH₂Br, —CCl₃, —CHCl₂, or —CH₂Cl; and

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, and glutamine. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In certain embodiments, R is selected from thegroup consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, tyrosine, isoleucine (including, R-isoleucine,S-isoleucine and RS-isoleucine), arginine, asparagine, and glutamine.

In certain embodiments, a compound of Formula (IA) is a compound inwhich X is imidazolyl or benzotriazolyl, and

(i) A and R are connected and form a side chain of proline, or

(ii) A is hydrogen, or a protecting group comprising phthalimidyl,tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, and R is selected fromthe group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, cysteine, serine, threonine, and glutamine. The side chainradicals may be unsubstituted or substituted with one or more of thefollowing: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl(e.g., a chloromethyl, a fluromethyl, etc.) or a protecting group (e.g.,Phth, Boc, Fmoc, Ddz, etc.).

In certain embodiments, a compound of Formula (IA) is a compound inwhich X is imidazolyl substituted with —CF₃, —CHF₂, —CH₂F, —CBr₃,—CHBr₂, —CH₂Br, —CCl₃, —CHC₂, or —CH₂Cl, and

(i) A and R are connected and form a side chain of proline, or

(ii) A is hydrogen, or a protecting group comprising phthalimidyl,tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, and R is selected fromthe group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, cysteine, serine, threonine, and glutamine. The side chainradicals may be unsubstituted or substituted with one or more of thefollowing: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl(e.g., a chloromethyl, a fluromethyl, etc.) or a protecting group (e.g.,Phth, Boc, Fmoc, Ddz, etc.).

In certain embodiments, a compound of Formula (IA) is a compound inwhich X is benzotriazolyl substituted with —CF₃, —CHF₂, —CH₂F, —CBr₃,—CHBr₂, —CH₂Br, —CCl₃, —CHCl₂, or —CH₂Cl, and

(i) A and R are connected and form a side chain of proline, or

(ii) A is hydrogen, or a protecting group comprising phthalimidyl,tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, and R is selected fromthe group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, threonine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, cysteine, serine, threonine, and glutamine. The side chainradicals may be unsubstituted or substituted with one or more of thefollowing: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl(e.g., a chloromethyl, a fluromethyl, etc.) or a protecting group (e.g.,Phth, Boc, Fmoc, Ddz, etc.).

In certain embodiments, compound of Formula (IA) is a compound, in whichR₁ and R are CH₂CH₂CH₂, and R₂ is tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, and X is is imidazolyl orbenzotriazolyl.

In certain embodiments, compound of Formula (IA) is a compound, in whichR₁ and R are CH₂CH₂CH₂, and R₂ is tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, and X is is imidazolyl orbenzotriazolyl.

The invention is also directed in part to compounds of Formula (IA):

wherein

X is imidazolyl or benzotriazolyl, and wherein

(i) A and R are connected and form a side chain of proline, or

(ii) A is hydrogen, or a protecting group comprising phthalimidyl,tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl; and R is selected fromthe group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, threonine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, cysteine, serine, threonine, and glutamine.

The invention is also directed in part to compounds of Formula (IA):

wherein

X is imidazolyl or benzotriazolyl, and wherein

(i) A and R are connected and form a side chain of proline, or

(ii) A is a protecting group selected from the group consisting ofphthalimidyl, tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl; and R is selected fromthe group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, threonine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, cysteine, serine, threonine, and glutamine.

The invention is also directed in part to compounds of Formula (IB):

wherein R₂ is a protecting group (e.g., tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl), and X is is imidazolylor benzotriazolyl. Compounds of Formula (IB) could be used in drugdiscovery, diagnosis, prevention and treatment of diseases, or asbuilding blocks for synthesis of peptidomimetic agents, e.g., for use indrug discovery, diagnosis, prevention and treatment of diseases.

In certain embodiments, the invention is directed to Phth-protectedcarbamoyl aza-imidazole derivatives and Phth-protected carbamoylaza-benzotriazole derivatives of unnatural amino acids, including, e.g.,aza-imidazole derivatives and Phth-protected carbamoyl aza-benzotriazolederivatives of β-amino acids (e.g., L-β-homotyrosine, β-alanine,L-β-homoasparagine, L-β-homoalanine, L-β-homophenylalanine,L-β-homoproline, L-β-holysine, L-β-homorarginine, L-β-proline, etc.),aliphatic amino acids (e.g., 6-aminohexanoic acid,2-amino-3-methoxybutanoic acid, 1-aminocyclopentane-1-carboxylic acid,2-(aminooxy)acetic acid, 6-aminohaxanoic acid,2-[2-(amino)-ethoxy]-ethoxy}acetic acid), β-cyclohexyl-L-alanine,6-aminohexanoic acid, L-α,β-diaminopropionic acid, L-propargylglycinel,L-α,β-diaminopropionic acid, α-aminoisobutyric acid,3-(2-pyridyl)-L-alanine, β-(3-pyridyl)-L-alanine,β-cyclopropyl-L-alanine, β-t-butyl-L-alanine,(2,4-dinitrophenyl))-L-α,β-diaminopropionic acid,(allyloxycarbonyl)-L-α,β-diaminopropionic acid, D-α,β-diaminopropionicacid, L-α,β-diaminopropionic acid,(N-γ-1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl)-L-α,γ-diaminobutyricacid, (N-γ-4-methyltrityl)-L-α,γ-diaminobutyric acid,L-α,γ-diaminobutyric acid, 4-fluoro-L-phenylglycine,5,5,5-trifluoro-DL-leucine, epsilon-aminohexanoic-OH,L-α-t-butylglycine, L-2-amino-3-(dimethylamino)propionic acid,L-2-aminocaproic acid, L-allylglycine, lysine azide,(Nδ-4-methyltrityl)-L-ornithine, Arg(Me)(Pbf)-OH, dimethyl-L-arginine(symmetrical and unsymmetrical), L-2-amino-3-guanidinopropionic acid,L-citrulline, F-acetyl-L-lysine, Lys(ivDde)-OH, Lys(Me)2-OH.HCl,Lys(Me3)-OHchloride, α-methyl-DL-glutamic acid, γ-carboxy-L-glutamicacid γ,γ-di-t-butyl ester, (N-γ-ethyl)-L-glutamine, 2,6-diaminopimelicacid, Glu(OAll)-OH, L-cysteic acid, α-methyl-DL-methionine,DL-buthionine, L-cysteic acid, L-selenomethionine,S-[2-(4-pyridyl)ethyl]-L-cysteine, S-[2-(4-pyridyl)ethyl]-L-cysteine,S-diphenylmethyl-L-cysteine, S-trityl-L-homocysteine,S-trityl-L-penicillamine, (Se-p-methoxybenzyl)-L-selenocysteine,β-hydroxyphenylalanine, 2-cyano-L-phenylalanine, L-thyroxine,O-methyl-L-tyrosine, β-methyl-DL-phenylalanine, 2-cyano-L-phenylalanine,L-thyroxine, O-methyl-L-tyrosine, β-methyl-DL-phenylalanine,2-cyano-L-phenylalanine, 3,4-dichloro-L-phenylalanine,3,4-difluoro-L-phenylalanine, 3,4-dihydroxy-L-phenylalanine,3,4-dihydroxy-phenylalanine, 3-amino-L-tyrosine, 3-chloro-L-tyrosine,3-fluoro-DL-tyrosine, 3-nitro-L-tyrosine, 4-amino-L-phenylalanine,4-aminomethyl-L-phenylalanine, 4-(phosphonomethyl)-phenylalanine,4-benzoyl-D-phenylalanine, 4-bis(2-chloroethyl)amino-L-phenylalanine,4-cyano-L-phenylalanine, 4-fluoro-L-phenylalanine,4-iodo-L-phenylalanine, DL-m-tyrosine, 2,6-dimethyl-tyrosine,L-homophenylalanine, O-methyl-L-tyrosine, Phe(4-guanidino)-OH,O-benzyl-L-phosphotyrosine, (2S,3R)-3-phenylpyrrolidine-2-carboxylicacid, (2S,4S)-4-phenyl-pyrrolidine-2-carboxylic acid,(2S,3aS,7aS)-Octahydro-1H-indole-2-carboxylic acid,(2S,3R)-3-phenylpyrrolidine-2-carboxylic acid,(2S,4R)-(−)-4-t-butoxypyrrolidine-2-carboxylic acid,trans-4-Fluoro-L-proline, (3S,4S)-4-amino-3-hydroxy-6-methylheptanoicacid, 4-amino-3-hydroxybutanoic acid, L-α-methylserine,(2S,3S)-2-amino-3-methoxybutanoic acid, Thr(β-D-GlcNAc(Ac)3)-OH,O-benzyl-L-phosphoserine, O-benzyl-D-phosphothreonine,O-benzyl-L-phosphothreonine, 4-methyl-DL-tryptophan,6-fluoro-DL-tryptophan, 6-methyl-DL-tryptophan, DL-7-azatryptophan,(R)-7-Azatryptophan, 5-benzyloxy-DL-tryptophan, 5-bromo-DL-tryptophan,5-chloro-DL-tryptophan, 5-fluoro-DL-tryptophan, 5-hydroxy-L-tryptophan,5-methoxy-L-tryptophan, 6-chloro-L-tryptophan, 6-methyl-DL-tryptophan,7-methyl-DL-tryptophan, DL-7-azatryptophan, 5-azido-pentanoic acid,2-Amino-N-(3-azidopropyl)-3-mercaptopropionamide,2-Amino-N-(3-azidopropyl)-3-mercaptopropionamide, Azidohomoalanine,L-propargylglycine.DCHA, azidolysine, p-azidophenylalanine,Azidohomoalanine, D-propargylglycine, L-propargylglycine, azidolysine,Tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl] amine, 2-(7′-octenyl)alanine, 2-(4′-pentenyl) alanine, 2-(4′-pentenyl)glycine, 2-(7′-octenyl)alanine, [5-((2-Aminoethyl)amino)naphthalene-1-sulfonic acid],L-glutamic acid-7-[2-(1-sulfonyl-5-naphthyl)-aminoethylamide],N-ε-(5-carboxyfluorescein)-L-lysine,N-ε-(5/6-carboxyfluorescein)-L-lysine,N-ε-(4,4-dimethylazobenzene-4′carbonyl)-L-lysine,Nε-2,4-dinitrophenyl-L-lysine,N-ε-[(7-methoxycoumarin-4-yl)-acetyl-L-lysine, glycosylated amino acids(e.g., Ser(β-D-GlcNAc(Ac)3)-OH, Thr(β-D-GlcNAc(Ac)3) -OH),3-azabicyclo[3.1.0]hexane-2-carboxylic acid, 4-amino-(1-carboxymethyl)piperidine, 4-phenylpiperidine-4-carboxylic acid,Na-methyl-N-im-trityl-L-histidine, Na-methyl-0-benzyl-L-serinedicyclohexylammonium salt,Nalpha-methyl-Nomega-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)-L-arginine,Nalpha-methyl-L-leucine, Nalpha-methyl-L-norvaline,Nalpha-methyl-L-phenylalanine, Nalpha-methyl-N-im-trityl-L-histidine,Nalpha-methyl-O-t-butyl-L-serine, Nalpha-methylglycine,21-amino-4,7,10,13,16,19-hexaoxaheneicosanoic acid,{2-[2-(amino)-ethoxy]-ethoxy}acetic acid, 6-Amino-4-oxahexanoic acid,5-Amino-3-Oxapentamoic Acid, NH-(PEG)10-CH2CH2COOH,NH-(PEG)12-CH2CH2COOH, 9-Amino-4; 7-Dioxanonanoic acid, 9-Amino-4;7-Dioxanonanoic acid, 12-amino-4,7,10-trioxadodecanoic acid,15-amino-4,7,10,13-tetraoxapentadecacanoic acid,18-amino-4,7,10,13,16-pentaoxaoctadecanoic acid,21-amino-4,7,10,13,16,19-hexaoxaheneicosanoic acid,NH-(PEG)8-CH2CH2COOH, 11-amino-3,6,9-trioxaundecanoic acid,N-(Fmoc-8-amino-3,6-dioxa-octyl)succinamic acid, —N-ε-acetyl-L-lysine,L-citrulline, Arg(Me)(Pbf)-OH, Nω,ω-dimethyl-L-arginine (assymetricaland symmetrical), Lys(Me)2-OH chloride, N-ε,ε-t-methyl-L-lysine,Lys(Me3)-OH chloride, O-benzyl-L-phosphoserine,O-benzyl-D-phosphothreonine, O-benzyl-L-phosphothreonine,O-benzyl-L-phosphotyrosine.

The invention is also directed to compounds of Formula (II):

wherein X is imidazolyl or benzotriazolyl; and

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, serine, and glutamine. The side chain radicals maybe unsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., N-Phth,N-Boc, N-Fmoc, N-Ddz, etc.). In certain embodiments, R is selected fromthe group consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, tyrosine, isoleucine (including, R-isoleucine,S-isoleucine and RS-isoleucine), arginine, asparagine, and glutamine.Compounds of Formula (II) could be used in drug discovery, diagnosis,prevention and treatment of diseases, or as building blocks forsynthesis of peptidomimetic agents, e.g., for use in drug discovery,diagnosis, prevention and treatment of diseases.

The invention is further directed to compounds of Formula (III):

wherein R is selected from the group consisting of side chain radicalsof aspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, threonine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, serine, and glutamine; and M is anoptional substituent selected from the group consisting of a halogen(Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH, —COH,methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., —CF₃, —CHF₂,—CH₂F, —CBr₃, —CHBr₂, —CH₂Br, —CCl₃, —CHCl₂, —CH₂Cl), —NH₂, or NH₃. Incertain embodiments, M is —CF₃. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In certain embodiments, R is selected from thegroup consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, threonine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, asparagine, andglutamine. Compounds of Formula (III) could be used in drug discovery,diagnosis, prevention and treatment of diseases, or as building blocksfor synthesis of peptidomimetic agents, e.g., for use in drug discovery,diagnosis, prevention and treatment of diseases.

The invention is also directed in part to the synthesis of compounds ofFormula (IA), (IB), (II) and (III), the synthesis comprising reacting aPhth-protected alkylhydrazine derivative with 1,1′-carbonyldiimidazole(CDI) or 1′-carbonyl-bis(3-ethylimidazolium) triflate (CBEIT).

The invention is further directed to compounds of Formula (IV):

wherein R is selected from the group consisting of side chain radicalsof aspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, threonine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, serine, and glutamine; and M is anoptional substituent selected from the group consisting of a halogen(Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH, —COH,methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., —CF₃, —CHF₂,—CH₂F, —CBr₃, —CHBr₂, —CH₂Br, —CCl₃, —CHCl₂, —CH₂Cl), —NH₂, or NH₃. Incertain embodiments, M is —CF₃. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In certain embodiments, R is selected from thegroup consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, tyrosine, isoleucine (including, R-isoleucine,S-isoleucine and RS-isoleucine), arginine, asparagine, and glutamine.Compounds of Formula (IV) could be used in drug discovery, diagnosis,prevention and treatment of diseases, or as building blocks forsynthesis of peptidomimetic agents, e.g., for use in drug discovery,diagnosis, prevention and treatment of diseases.

The invention is also directed in part to the synthesis of the compoundsof Formula (IV), the synthesis comprising reacting an acid chloride withbenzotriazole:

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (V):

wherein

is (i) at the N-terminus and/or the C-terminus and/or (ii) adjacent tothe N-terminus and/or the C-terminus of the compound of Formula (V),wherein

B is selected from the group consisting of hydrogen, —NH₂, —NNH₂,—CONH₂, —COOR₃, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D is selected from the group consisting of —OR₄, —OH, —NH₂, —NNH₂,—NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60-mer aza peptide, a 2 to 60-mer azatide;

R₃ and R₄ is each independently selected from the group consisting ofC₁-C₆ alkyl (e.g., methyl), methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.);

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, threonine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, serine, threonine, cysteine, andglutamine. The side chain radical of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, serine, threonine,cysteine and glutamine may be unsubstituted or substituted with one ormore of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.). In certain embodiments,compounds of Formula (V) are peptidomimetic analogues of compounds ofFormula (VI):

or a pharmaceutically acceptable salt thereof; wherein

B is selected from the group consisting of hydrogen, —NH₂, —NNH₂,—CONH₂, —COOR₃, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D is selected from the group consisting of —OR₄, —OH, —NH₂, —NNH₂,—NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60-mer aza peptide, a 2 to 60-mer azatide;

R₃ and R₄ is each independently selected from the group consisting ofC₁-C₆ alkyl (e.g., methyl), methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.);

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, serine, threonine, cysteine, and glutamine. Theside chain radical of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, threonine,cysteine and glutamine may be unsubstituted or substituted with one ormore of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.); and compounds of Formula (V)are more resistant to hydrolysis and/or enzymatic degradation thancompounds of Formula (VI). In some of these embodiments, compounds ofFormula (V) are more potent than compounds of Formula (VI), e.g., due toa better fit into a biological receptor.

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (V):

wherein

is at or adjacent to a cleavage and/or a hydrolysis site of the compoundof Formula (V), wherein

B is selected from the group consisting of hydrogen, —NH₂, —NNH₂,—CONH₂, —COOR₃, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D is selected from the group consisting of —OR₄, —OH, —NH₂, —NNH₂,—NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60-mer aza peptide, a 2 to 60-mer azatide;

R₃ and R₄ is each independently selected from the group consisting ofC₁-C₆ alkyl (e.g., methyl), methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.);

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, serine, threonine, cysteine, and glutamine. Theside chain radical of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, threonine,cysteine and glutamine may be unsubstituted or substituted with one ormore of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.). In certain embodiments,compounds of Formula (V) are peptidomimetic analogues of compounds ofFormula (VI):

or a pharmaceutically acceptable salt thereof; wherein

B is selected from the group consisting of hydrogen, —NH₂, —NNH₂,—CONH₂, —COOR₃, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D is selected from the group consisting of —OR₄, —OH, —NH₂, —NNH₂,—NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60-mer aza peptide, a 2 to 60-mer azatide;

R₃ and R₄ is each independently selected from the group consisting ofC₁-C₆ alkyl (e.g., methyl), methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.);

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, serine, threonine, cysteine, and glutamine. Theside chain radical of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, threonine,cysteine and glutamine may be unsubstituted or substituted with one ormore of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.); and compounds (V) are moreresistant to hydrolysis and/or enzymatic degradation than compounds ofFormula (VI). In some of these embodiments, compounds of Formula (V) aremore potent than compounds of Formula (VI), e.g., due to a better fitinto a biological receptor.

In certain embodiments, B of compounds of Formula (V) and (VI) is eachindependently selected from the group consisting of hydrogen, —NH₂,—NNH₂, —CONH₂, —COOR₃, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, an aminoacid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D of compounds of Formula (V) and (VI) is each independently selectedfrom the group consisting of —OR₄, —NH₂, —NNH₂, —NHCOCH₃, —NHCH₃,—N(CH₃)2, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

R₃ and R₄ of compounds of Formula (V) and (VI) is each independentlyselected from the group consisting of C₁-C₆ alkyl (e.g., methyl),methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., a chloromethyl, afluromethyl, etc.) or a protecting group (e.g., Phth, Boc, Fmoc, Ddz,etc.); and

R of compounds of Formula (V) and (VI) is each independently selectedfrom the group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, serine, threoinine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, and glutamine. The side chain radical of asparticacid, phenylalanine, alanine, histidine, glutamic acid, tryptophan,valine, leucine, lysine, methionine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, may be unsubstituted or substituted with one or more of thefollowing: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl(e.g., a chloromethyl, a fluromethyl, etc.) or a protecting group (e.g.,Phth, Boc, Fmoc, Ddz, etc.).

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (V):

wherein

is adjacent to the N-terminus and/or the C-terminus of the compound ofFormula (V), wherein

B is selected from the group consisting of hydrogen, —NH₂, —NNH₂,—CONH₂, —COOR₃, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D is selected from the group consisting of —OR₄, —OH, —NH₂, —NNH₂,—NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60-mer aza peptide, a 2 to 60-mer azatide;

R₃ and R₄ is each independently selected from the group consisting ofC₁-C₆ alkyl (e.g., methyl), methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.);

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, serine, threonine, cysteine, and glutamine. Theside chain radical of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, threonine,cysteine and glutamine may be unsubstituted or substituted with one ormore of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.). In certain embodiments,compounds of Formula (V) are peptidomimetic analogues of compounds ofFormula (VI):

or a pharmaceutically acceptable salt thereof; wherein

B is selected from the group consisting of hydrogen, —NH₂, —NNH₂,—CONH₂, —COOR₃, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D is selected from the group consisting of —OR₄, —OH, —NH₂, —NNH₂,—NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄haloalkyl, an amino acid, an aza amino acid, a 2 to 60-mer peptide, a 2to 60-mer aza peptide, a 2 to 60-mer azatide;

R₃ and R₄ is each independently selected from the group consisting ofC₁-C₆ alkyl (e.g., methyl), methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.);

R is selected from the group consisting of side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, threonine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, serine, threonine, cysteine, andglutamine. The side chain radical of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, serine, threonine,cysteine and glutamine may be unsubstituted or substituted with one ormore of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COGH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.); and compounds (V) are moreresistant to hydrolysis and/or enzymatic degradation than compounds ofFormula (VI). In some of these embodiments, compounds of Formula (V) aremore potent than compounds of Formula (VI), e.g., due to a better fitinto a biological receptor.

Compounds of Formula (V) could be used, e.g., in drug discovery,diagnosis, prevention and treatment of diseases.

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (VII):

or a pharmaceutically acceptable salt thereof,

wherein E is hydrogen, —NH₂, —NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, —OH, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60 mer aza peptide, or a 2 to 60-mer azatide;

G is —OH, —NH₂, —NNH₂, —NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH,—COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a2 to 60-mer peptide, a 2 to 60-mer aza peptide, or a 2 to 60-merazatide;

R is selected from the group consisting of unsubstituted and substitutedside chain radicals of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, and glutamine,

wherein

is at or adjacent to a cleavage and/or a hydrolysis site and/or at theN-terminus and/or the C-terminus of the compound of Formula (VII). Theside chain radicals may be unsubstituted or substituted with one or moreof the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, a C₁-C₆ haloalkyl(e.g., a chloromethyl, a fluromethyl, etc.) or a protecting group (e.g.,Phth, Boc, Fmoc, Ddz, etc.). In some embodiments, R is methyl serine,methyl threonine or methyl cysteine. Compounds of Formula (VII) are moreresistant to hydrolysis and/or enzymatic degradation than compounds thatcontain

instead of

but are otherwise identical to the compounds of Formula (VII).

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (VII):

or a pharmaceutically acceptable salt thereof,

wherein E is hydrogen, —NH₂, —NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, —OH, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60 mer aza peptide, or a 2 to 60-mer azatide;

G is —OH, —NH₂, —NNH₂, —NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH,—COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a2 to 60-mer peptide, a 2 to 60-mer aza peptide, or a 2 to 60-merazatide;

R is selected from the group consisting of unsubstituted and substitutedside chain radicals of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, and glutamine,

wherein

is adjacent the N-terminus and/or the C-terminus of the compound ofFormula (VII). The side chain radicals may be unsubstituted orsubstituted with one or more of the following: a halogen (Cl, F, or Br),a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl,a C₁-C₆ haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or aprotecting group (e.g., Phth, Boc, Fmoc, Ddz, etc.). In someembodiments, R is methyl serine, methyl threonine or methyl cysteine.Compounds of Formula (VII) are more resistant to hydrolysis and/orenzymatic degradation than compounds that contain

instead of

but are otherwise identical to the compounds of Formula (VII).

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (VII):

or a pharmaceutically acceptable salt thereof,

wherein E is hydrogen, —NH₂, —NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, —OH, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60 mer aza peptide, or a 2 to 60-mer azatide;

G is —OH, —NH₂, —NNH₂, —NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH,—COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a2 to 60-mer peptide, a 2 to 60-mer aza peptide, or a 2 to 60-merazatide;

R is selected from the group consisting of unsubstituted and substitutedside chain radicals of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine arginine, glycine, asparagine, serine, and glutamine,

wherein

is adjacent the N-terminus of the compound of Formula (VII). The sidechain radicals may be unsubstituted or substituted with one or more ofthe following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In some embodiments, R is methyl serine, methylthreonine or methyl cysteine. Compounds of Formula (VII) are moreresistant to hydrolysis and/or enzymatic degradation than compounds thatcontain

instead of

but are otherwise identical to the compounds of Formula (VII).

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (VII):

or a pharmaceutically acceptable salt thereof,

wherein E is hydrogen, —NH₂, —NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, —OH, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60 mer aza peptide, or a 2 to 60-mer azatide;

G is —OH, —NH₂, —NNH₂, —NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH,—COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a2 to 60-mer peptide, a 2 to 60-mer aza peptide, or a 2 to 60-merazatide;

R is selected from the group consisting of unsubstituted and substitutedside chain radicals of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, and glutamine,wherein

is adjacent the C-terminus of the compound of Formula (VII). The sidechain radicals may be unsubstituted or substituted with one or more ofthe following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In some embodiments, R is methyl serine, methylthreonine or methyl cysteine. Compounds of Formula (VII) are moreresistant to hydrolysis and/or enzymatic degradation than compounds thatcontain

instead of

but are otherwise identical to the compounds of Formula (VII).

The invention is further directed to the use of compounds of Formulas(I)-(IV) in the preparation of compounds of Formula (VII):

or a pharmaceutically acceptable salt thereof, wherein

is at or adjacent to a cleavage and/or a hydrolysis site and/or at theN-terminus and/or the C-terminus of the compound of Formula (VII),

wherein E is hydrogen, —NH₂, —NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, —OH, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60 mer aza peptide, or a 2 to 60-mer azatide;

G is —OH, —NH₂, —NNH₂, —NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH,—COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a2 to 60-mer peptide, a 2 to 60-mer aza peptide, or a 2 to 60-merazatide;

R is selected from the group consisting of unsubstituted and substitutedside chain radicals of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, and glutamine.The side chain radicals may be unsubstituted or substituted with one ormore of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, a C₁-C₆ haloalkyl(e.g., a chloromethyl, a fluromethyl, etc.) or a protecting group (e.g.,Phth, Boc, Fmoc, Ddz, etc.). In some embodiments, R is methyl serine,methyl threonine or methyl cysteine. In certain embodiments, compoundsof Formula (VII) are peptidomimetic analogues of compounds of Formula(VIII)

or a pharmaceutically acceptable salt thereof,

wherein E is hydrogen, —NH₂, —NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, —OH, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60 mer aza peptide, or a 2 to 60-mer azatide;

G is —OH, —NH₂, —NNH₂, —NHCOCH₃, —NHCH₃, —N(CH₃)2, —CONH₂, —COOH, —COH,—COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a2 to 60-mer peptide, a 2 to 60-mer aza peptide, or a 2 to 60-merazatide;

R is selected from the group consisting of unsubstituted and substitutedside chain radicals of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, glycine, asparagine, serine, and glutamine;

wherein the side chain radicals may be unsubstituted or substituted withone or more of the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl(e.g., methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.);

wherein compounds of Formula (VII) are more resistant to hydrolysisand/or enzymatic degradation than compounds of Formula (VIII). In someof these embodiments, compounds of Formula (VII) are more potent thancompounds of Formula (VIII), e.g., due to a better fit into a biologicalreceptor. In some embodiments, R of each compound is selected from agroup consisting of methyl serine, methyl threonine and methyl cysteine.

The invention is also directed to compounds of Formula (V) and compoundsof Formula (VII). Compounds of Formula (V) and (VII) could be used,e.g., in drug discovery, diagnosis, prevention and treatment ofdiseases.

The replacement of

at the N-terminus and/or the C-terminus and/or at or adjacent to acleavage and/or a hydrolysis site of compounds of Formula (VI) andFormula (VIII) with

results in a loss of asymmetry associated with

in compounds of Formula (VI) and Formula (VIII). Because of the changein configuration, compounds of Formula (V) and Formula (VII) may betherapeutically effective, whereas compounds of Formula (VI) and Formula(VIII) are not, and/or compounds of Formula (V) and Formula (VII) mayhave a greater bioavailability (e.g., oral and/or transdermal and/orintranasal) than compounds of Formula (VI) and compounds of Formula(VIII), and/or compounds of Formula (V) and Formula (VII) may have an invivo half-life greater than (e.g., more than twice, three times, fivetimes or ten times) the in vivo half-life of the compounds of Formula(VI) and (VIII), while maintaining therapeutic efficacy, and/orcompounds of Formula (V) and Formula (VII) may have a longer duration oftherapeutic activity than compounds of Formula (VI) and Formula (VIII),and/or compounds of Formula (V) and Formula (VII) may have a greateraffinity to a biological receptor than compounds of Formula (VI) andFormula (VIII), and/or compounds of Formula (V) and Formula (VII) mayact as agonists at a biological receptor, whereas compounds of Formula(VI) and Formula (VIII) act as antagonists of the biological receptor,and/or compounds of Formula (V) and Formula (VII) may act as antagonistsat the biological receptor, whereas compounds of Formula (VI) andFormula (VIII) act as agonists at the biological receptor.

Therefore, in certain embodiments, the invention is directed to acompound of Formula (V), which is a peptidomimetic analogue of acompound of Formula (VI) and is more resistant to enzymatic degradation(e.g., peptidase degradation) than the compound of Formula (VI). In someof these embodiments, the compound of Formula (V) is at least 1.5, 2, 3,4 or 5 more resistant to enzymatic degradation than the compound ofFormula (VI). In certain embodiments, the compounds of Formula (V) isabout 10, 12, 14, 16, 18, or 20 times more resistant to enzymaticdegradation than the compound of Formula (VI).

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI) and has a longer in vitro and/or in vivo half-life than thecompound of Formula (VI). In some of these embodiments the in vitroand/or in vivo half-life of the compound of Formula (IX) is at least1.5, 2, 3, 4 or 5 times longer than the corresponding half-life of thecompound of Formula (VI). In certain embodiments, the compound ofFormula (V) has an in vivo half-life greater than (e.g., twice, threetimes, five times or ten times) of the in vivo half-life of the compoundof Formula (VI) and maintains therapeutic efficacy of the compound ofFormula (VI).

In certain embodiments, the in vivo half-life of the compound of Formula(V) may, e.g., be from about 1 minute to about 30 days, and the in vivohalf-life of the compound of Formula (VI) may, e.g., be from 5 secondsto about 10 days. In certain embodiments, the in vivo half-life of thecompound of Formula (V) is from about 1 minute to about 72 hours, andthe in vivo half-life of the compound of Formula (VI) is from about 10seconds to 24 hours.

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI) and has a greater biovailability than the compound of Formula (VI).The bio-availability (e.g., oral) of compound of Formula (V) may, e.g.,be at least 10, 20, 30, 40, 50, 60, or 70% better than thebio-availabilty of the compound of Formula (VI).

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI) and has greater blood brain barrier (BBB) permeability than acompound of Formula (VI). The BBB permeability of compound of Formula(V) may, e.g., be at least 10, 20, 30, 40, 50, 60, or 70% better thanthe BBB permeability of the compound of Formula (VI).

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI) and has a greater therapeutic efficacy than the compound of Formula(VI).

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI) and has a longer duration of therapeutic activity than the compoundof Formula (VI). In some of these embodiments, the duration of action ofthe compound of Formula (V) is at least double, triple, or quadriple theduration of action of the compound of Formula (V). In certainembodiments, the duration of action of the compound of Formula (V) isfrom about 5 minutes to about 24 hours, from about 10 minutes to about22 hours, from about 15 minutes to about 20 hours, from about 30 minutesto about 20 hours, from about 45 minutes to about 20 hours, from about 1hour to about 20 hours, from about 2 hours to about 18 hours, from about2 hours to about 16 hours, from about 2 hours to about 14 hours, fromabout 2 hours to about 12 hours, from about 2 hours to about 10 hours,from about 3 hours to about 12 hours, from about 4 hours to about 12hours, from about 4 hours to about 10 hours, or from about 4 hours toabout 8 hours.

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI) and has a greater affinity for a biological receptor than thecompound of Formula (VI). For example, the binding affinity (KD) of thecompound of Formula (V) to the biological receptor may, e.g., be from1×10⁻¹² M to 1×10⁻⁴ M, and is at least 1.5, 2, 3, 4 or 5 times strongerthan the affinity of the compound of formula (VI) to the biologicalreceptor. The biological receptor, may e.g., be an ANP receptor, an AVPreceptor, a B2 receptor, a BNP receptor, a CCK receptor, a CALCreceptor, a CALC receptor and RAMPs, a CRH receptor, a CD 36 receptor, aCD110 receptor, a CXCR4 receptor, an EPO receptor, a FGF receptor, aET-B receptor, a GCG receptor, a GH receptor, a GNRH receptor, a GnRH Rreceptor, a GPL-1 receptor, a GPL-2 receptor, a GHS receptor, a GPR54, aGuanylate cyclase-C, a IL2 receptor, a IGF-1 receptor, a PGE2 receptor,a NGF receptor, a NMDA receptor, a NOD protein receptor, a NPY receptor,a MC receptor, a M1 receptor, a NTS1 receptor, a NK receptor, a PTHreceptor, a Delta opioid receptor, a Kappa opioid receptor, a Mu opioidreceptor, an ORL1 receptor, an OGF receptor, an OT receptor, a PARreceptor, a SCT receptor, a SST receptor, a SST receptor and Dopamine D2receptor, a TRH receptor, a VPAC receptor, or RAMPs receptor.

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI) and binds a biological receptor slower but stronger than thecompound of Formula (VI). The compound of Formula (V), preferably, has abetter KD affinity than the parent peptide.

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI), wherein the compound of Formula (V) is an agonist at a biologicalreceptor and the compound of Formula (VI) is an antagonist at thebiological receptor. The biological receptor, may e.g., be an ANPreceptor, an AVP receptor, a B2 receptor, a BNP receptor, a CCKreceptor, a CALC receptor, a CALC receptor and RAMPs, a CRH receptor, aCD 36 receptor, a CD110 receptor, a CXCR4 receptor, an EPO receptor, aFGF receptor, a ET-B receptor, a GCG receptor, a GH receptor, a GNRHreceptor, a GnRH R receptor, a GPL-1 receptor, a GPL-2 receptor, a GHSreceptor, a GPR54, a Guanylate cyclase-C, a TL2 receptor, a IGF-1receptor, a PGE2 receptor, a NGF receptor, a NMDA receptor, a NODprotein receptor, a NPY receptor, a MC receptor, a M1 receptor, a NTS1receptor, a NK receptor, a PTH receptor, a Delta opioid receptor, aKappa opioid receptor, a Mu opioid receptor, an ORL1 receptor, an OGFreceptor, an OT receptor, a PAR receptor, a SCT receptor, a SSTreceptor, a SST receptor and Dopamine D2 receptor, a TRH receptor, aVPAC receptor, or RAMPs receptor.

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI), wherein the compound of Formula (V) is an antagonist at abiological receptor and the compound of Formula (VI) is an agonist atthe biological receptor. The biological receptor may, e.g., be an ANPreceptor, an AT receptor, a B-cell activating factor, a B2 receptor, aBB2 receptor, N-Cadherin, calcium channel, a CCRP5 receptor, a CALCreceptor and RAMPs, a CD4 receptor, a C5a receptor, a CD29 receptor, aCXCR4 receptor, a GCG receptor, a Erb-3 receptor, a GnRH R, a GP IIbIIIa receptor, an integrin alpha-5/beta-3, an integrin alpha-4/beta-1,an NMDA receptor, a Nicotinic ACH receptor, an OT receptor, a PTHreceptor, a SST receptor, a TACl receptor, a TAC2 receptor, a TBXA2receptor, a VEGF receptor, a VE-Cadherin receptor, or a zonulinreceptor.

In certain embodiments, the invention is directed to a compound ofFormula (V), which is a peptidomimetic analogue of a compound of Formula(VI), wherein the compound of Formula (VI) is not therapeuticallyeffective, and the compound of Formula (V) is therapeutically effective.

The invention is also directed in part to a method of preparing acompound of Formula (V), the method comprising activating a compound ofFormula (IA), Formula (IB), Formula (II), Formula (III) or Formula (IV)to form an activated compound of Formula (IA), Formula (IB), Formula(II), Formula (III), or Formula (IV), and coupling the activatedcompound of Formula (IA), Formula (IB), Formula (II), Formula (III), orFormula (IV) with N-terminal of an amino acid, N-terminal of an aza-minoacid, provided that, if a side chain of the amino acid or aza-amino acidcontains a group selected from amino, amide, guanidino N, carboxyl,sulfhydryl, carboxyl, hydroxyl, indole, imidazole phenol, the group isprotected with a protecting group selected from tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl, or2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, phthalimide,carboxybenzyl, 2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl,trityl or triphenylmethyl, t-butyl ester, t-butyl ether, s-t-butylether, allyloxycarbonyl, methoxytrimethylbenzene sulfonyl,4,4-dimethyloxybenzhydryl, 2,2,5,7,8-pentamethyl-chroman-6-sulfonylchloride, 2,4,6-trimethoxybenzyl, allyl ester acetamidomethyl, and thelike to form a protected compound of Formula (V), and deprotecting theprotected compound of Formula (V), e.g., with hydrazine, piperadine,TFA, acetic acid, thioanisole, EDT, anisole, etc., to form the compoundof Formula (V).

In certain embodiments, the compound of Formula (IA), Formula (IB),Formula (II), or Formula (III) is phthalimide-protected carbamoylimidazole and is activated by Mel.

In certain embodiments, the compound of Formula (IA), Formula (IB),Formula (II) or Formula (III) is phthalimide-protected carbamoylbenzotriazole and is activated by DIPEA in acetonitrile.

The invention is also directed in part to a method of preparing acompound of Formula (V), the method comprising coupling a compound ofFormula (IA), (IB), (II), (III), or (IV) with an aza-amino acid to forma protected di-azatide, and deprotecting the protected di-azatide, e.g.,with hydrazine, TFA, acetic acid, thioanisole, EDT, anisole, a mixtureof any of the foregoing, or another de-protecting compound, to form acompound of Formula (VI).

The invention is further directed in part to a solution phase synthesisof the compounds of Formula (V), the solution phase synthesis comprisingconverting a compound of Formula (IA), I(B), (II), (III), or (IV) to anamide of the compound of Formula (IA), I(B), (II), (III), or (IV),deprotectecting the amide of the compound of Formula (IA), I(B), (II),(III), or (IV), and coupling the deprotected amide of Formula (IA),I(B), (II), (III), or (IV) with an additional compound of Formula (IA),I(B), (II), (III), or (IV), or a protected amino acid, or a protectedaza-amino acid to form a protected azapeptide, and deprotecting theprotected azapeptide to provide a compound of Formula (V).

The invention is further directed in part to a solid phase synthesis ofthe compounds of Formula (V), the solid phase synthesis comprisingcoupling a protected compound of Formula (IA), I(B), (II), (III), or(IV) to a support, deptrotecting the protected compound of Formula (IA),I(B), (II), (III), or (IV), and coupling the deprotected compound ofFormula (IA), I(B), (II), (III), or (IV) to an additional protectedcompound of Formula (IA), I(B), (II), (III), or (IV), an additionalprotected amino acid, or an additional protected aza-amino acid to forma protected peptide, and deprotecting and cleaving the protected peptideto provide a compound of Formula (V).

The invention is also directed in part to a process of preparing acompound of Formula (V) comprising cleaving a peptide at its N-terminusand/or C-terminus, and coupling the cleaved peptide with a compound ofFormula (IA), I(B), (II), (III), or (IV) to form a compound of Formula(V). In certain embodiments, the compound of Formula (IA), I(B), (II),(III), or (IV) is activated prior to the coupling with the cleavedpeptide.

The invention is also directed in part to a process of preparing acompound of Formula (V) comprising cleaving a peptide at its cleavagesite to form two smaller peptides, replacing the last amino acid of atleast one of the smaller peptides with an aza-amino acid to form anazapeptide, and conjugating the azapeptide with the remaining smallerpeptide to provide a compound of Formula (V).

The invention is also directed in part to a process of preparing acompound of Formula (V) comprising hydrolizing a peptide at its cleavagesite, and reacting the cleaved peptide with a compound of Formula (IA),I(B), (II), (III), or (IV) to provide a compound of Formula (V).

The processes for preparing compounds of Formula (VII) are identical tothe processes of preparing compounds of Formula (V).

The invention is also directed in part to a method of synthesis of anazapeptide comprising coupling a compound of Formula (IA), I(B), (II),(III), or (IV) to an amino acid or an aza-amino acid, wherein theazapeptide is a compound of Formula (V):

or a pharmaceutically acceptable salt thereof, wherein

B is selected from the group consisting of hydrogen, —NH₂, —NNH₂,—CONH₂, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, an aminoacid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide;

D is selected from the group consisting of —OH, —NH2, —NNH2, —CONH2,—COOH, —COH, —COC1-C4 alkyl, —COC1-C4 haloalkyl, an amino acid, an azaamino acid, a 2 to 60-mer peptide, a 2 to 60-mer aza peptide, a 2 to60-mer azatide; and

R is selected from the group consisting of unsubstituted and substitutedside chain radicals of aspartic acid, phenylalanine, alanine, histidine,glutamic acid, tryptophan, valine, leucine, lysine, methionine,tyrosine, isoleucine, arginine, glycine, asparagine, serine, andglutamine. The coupling can be either during a solid phase peptidesynthesis or a liquid phase peptide synthesis. The method may compriseactivating the compound Formula (IA), I(B), (II), (III), or (IV) priorto the coupling. The activating may be with, e.g., with Me or DIPEA. Incertain embodiments, the compound of Formula (V) is produced in a yieldof at least about 50% (e.g., about 55%, about 60%, about 65%, about 70%,about 75%, about 80%, about 85%, about 90%, about 95%, or about 97%) andthe compound of Formula (IA) is:

wherein

X is imidazolyl or benzotriazolyl, and wherein

(i) A and R are connected and form a side chain of proline, or

(ii) A is hydrogen, or a protecting group comprising phthalimidyl,tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl; and R is selected fromthe group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, tyrosine, isoleucine, arginine, glycine,asparagine, serine, cysteine, serine, threonine, and glutamine.

The invention is further directed in part to a method of azapeptidesynthesis comprising reacting (i) an imidazole derivative comprising anaza-amino acid, wherein the aza-amino acid is covalently bound(conjugated) to a protecting group at its N-terminus and to imidazole atits C-terminus and is selected from the group consisting of aza-glycine,aza-alanine, aza-valine, aza-leucine, aza-isoleucine, aza-proline,aza-phenylalanine, aza-tyrosine, aza-tryptophan, aza-aspartic acid,aza-glutamic acid, aza-asparagine, aza-glutamine, aza-histidine,aza-lysine, and aza-arginine with (ii) an aza-amino acid, an amino acid,or a peptide to form the azapeptide, wherein the imidazole derivative isa compound of Formula (IA), (IIB) or (III), and the azapeptide is acompound of formula (V) or (VII).

The invention is also directed in part to a method of an azapeptidesynthesis comprising reacting (i) an imidazole derivative comprising anaza-amino acid, wherein the aza-amino acid is covalently bound(conjugated) to a protecting group at its N-terminus and to imidazole atits C-terminus and is selected from the group consisting of aza-glycine,aza-alanine, aza-valine, aza-leucine, aza-isoleucine, aza-proline,aza-phenylalanine, aza-tyrosine, aza-tryptophan, aza-aspartic acid,aza-glutamic acid, aza-aspargine, aza-glutamine, aza-histidine,aza-lysine, and aza-arginine with (ii) a peptide to form the azapeptide,wherein the imidazole derivative is a compound of Formula (IA), (IB),(II) or (III), and the azapeptide is a compound of formula (V) or (VII).

In addition, the invention is directed in part to a method of anazapeptide synthesis comprising reacting (i) a benzotriazole derivativecomprising an aza-amino acid covalently bound (conjugated) to a firstprotecting group at its N-terminus and to benzotriazole at itsC-terminus, wherein the aza-amino acid is selected from the groupconsisting of aza-glycine, aza-alanine, aza-valine, aza-leucine,aza-isoleucine, aza-proline, aza-phenylalanine, aza-tyrosine,aza-tryptophan, aza-aspartic acid, aza-glutamic acid, aza-aspargine,aza-aspartic acid, aza-glutamine, aza-histidine, aza-lysine, andaza-arginine with (ii) a an aza-amino acid, an an amino acid, or apeptide to form the azapeptide, wherein the benzotriazole derivative isa compound of Formula (IV), and the azapeptide is a compound of formula(V) or (VII), provided that if a side chain of the aza-amino acidcomprises a group selected from amino, amide, guanidino N, carboxyl,sulfhydryl, carboxyl, hydroxyl, indole, imidazole phenol, the group isprotected by a second protecting group. The first and the secondprotecting group may independently be, e.g., a protecting group selectedfrom tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, or2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, phthalimide,carboxybenzyl, 2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl,trityl or triphenylmethyl, t-butyl ester, t-butyl ether, s-t-butylether, allyloxycarbonyl, methoxytrimethylbenzene sulfonyl,4,4-dimethyloxybenzhydryl, 2,2,5,7,8-pentamethyl-chroman-6-sulfonylchloride, 2,4,6-trimethoxybenzyl, allyl ester, acetamidomethyl, and thelike. In certain embodiments, the first and second protecting group iseach independently selected from the group consisting of betert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, or2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl, phthalimide, orcarboxybenzyl.

The invention is also directed to a method of azapeptide synthesiscomprising reacting (i) a benzotriazole derivative comprising anaza-amino acid, wherein the aza-amino acid is covalently bound(conjugated) to a first protecting group at its N-terminus and tobenzotriazole at its C-terminus and is selected from the groupconsisting of aza-glycine, aza-alanine, aza-valine, aza-leucine,aza-isoleucine, aza-proline, aza-phenylalanine, aza-tyrosine,aza-tryptophan, aza-aspartic acid, aza-glutamic acid, aza-aspargine,aza-aspartic acid, aza-glutamine, aza-histidine, aza-lysine, andaza-arginine with (ii) a peptide to form the azapeptide, wherein thebenzotriazole derivative is a compound of Formula (IV), and theazapeptide is a compound of formula (V) or (VII), provided that if aside chain of the aza-amino acid comprises a group selected from amino,amide, guanidino N, carboxyl, sulfhydryl, carboxyl, hydroxyl, indole,imidazole phenol, the group is protected by a second protecting group.The first and the second protecting group may independently be, e.g., aprotecting group selected from tert-butoxycarbonyl (Boc),9-fluorenylmethoxycarbonyl (Fmoc), or2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl (Ddz), phthalimide (Phth),carboxybenzyl (Cbz), 2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl(Pbf), trityl or triphenylmethyl (Trt), t-butyl ester (OtBu), t-butylether (tBu), s-t-butyl ether, allyloxycarbonyl (Aloc),methoxytrimethylbenzene sulfonyl (Mtr), 4,4-dimethyloxybenzhydryl (Mbh),2,2,5,7,8-pentamethyl-chroman-6-sulfonyl chloride (Pmc),2,4,6-trimethoxybenzyl (Tmob), allyl ester (OAI), acetamidomethyl (Acm),and the like. In certain embodiments, the first and second protectinggroup is each independently selected from the group consisting of betert-butoxycarbonyl (Boc), 9-fluorenylmethoxycarbonyl (Fmoc), or2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl (Ddz), phthalimide (Phth),or carboxybenzyl (Cbz).

The compounds of Formula (IA), (IB), (II), (III), or (IV) and process ofthe invention allow, e.g., for preparation of a compound of Formula (V)in yields of at least about 50% (e.g., from about 55% to about 99%, fromabout 60% to about 95%, or from about 65% to about 95%). Thus, the yieldmay, e.g., be about 55%, about 60%, about 65%, about 75%, about 80%,about 85%, about 90%, about 95%, about 97%, or about 99%. In certainembodiments, the yield is greater than 85%.

The invention is further directed in part to prolonging an in vitroand/or in vivo half-life of a peptide, comprising synthesizing apeptidomimetic analogue of the peptide with the compounds of Formula(I), (II), (III) or (IV), the peptidomimetic analogue containing an azaamino acid instead of amino acid at the N-terminus of the peptide, butis otherwise identical to the peptide. In certain embodiments, thepeptidomimetic analogue is a compound of Formula (V), (VII), (IX), or(X), and is synthesized by liqid phase or solid phase chemistry.

The invention is further directed in part to prolonging an in vitroand/or in vivo half-life of a peptide, comprising synthesizing apeptidomimetic analogue of the peptide with the compounds of Formula(I), (II), (III) or (IV), the peptidomimetic analogue containing an azaamino acid instead of amino acid at a position adjacent to theN-terminus of the peptide, but is otherwise identical to the peptide. Incertain embodiments, the peptidomimetic analogue is a compound ofFormula (V), (VII), (IX), or (X), and is synthesized by liqid phase orsolid phase chemistry.

The invention is further directed in part to the use of compounds ofFormula (V), (VII), (IX), or (X) in prevention, diagnosis and treatmentof medical conditions, including, e.g., cardiovascular disorders, CNSdisorders, neurodegenerative disorders, immune system disorders,metabolic disorders, fertility, dental conditions, pain, inflammation,dermatological conditions, blood disorders, infection, eye disorders,gynecologic disorders, urologic disorders, bone and connective tissuedisorders, respiratory disorders, gastrointestinal disorders, disordersof endocrine system, and cancer.

The methods of diagnosing, prevention and treatment of medicalconditions in accordance with the present invention compriseadministering a therapeutically effective amount of a compound ofFormula (V) or Formula (VII) to a subject in need thereof at specifictimes in a pharmaceutically acceptable formulation.

In certain embodiments, the method of treating a disorder comprisingco-administering a compound of Formula (V) or (VII) along with acompound of Formula (VI) or (VIII). In certain embodiments, the compoundof Formula (V) or Formula (VII) acts as a competitive inhibitor of thecompound of Formula (VI) or Formula (VIII). In certain embodiments, theadministration of the compound of Formula (V) or (VII) results in higherplasma concentrations of the compound of Formula (VI) or (VIII).

In certain embodiments, the invention is directed to a method ofprolonging effects of a 2 to 50 amino acid peptide comprisingadministering, before, after or concurrently with the peptide, anaza-analogue of the amino acid peptide, the analogue differing frompeptide in that at least one of the amino acids of the peptide isreplaced with a corresponding aza-amino acid.

The invention is further directed to a pharmaceutically acceptableformulation comprising a therapeutically effective amount of a compoundof Formula (V) or Formula (VII) and one or more pharmaceuticallyacceptable excipient(s). The pharmaceutically acceptable excipients aredescribed in the the Handbook of Pharmaceutical Excipients,Pharmaceutical Press and American Pharmacists Association, sixth ed.,(2009), incorporated by reference herein, for all purposes.

The invention is further directed to a diagnostic formulation comprisinga compound of Formula (V) or Formula (VII).

Definitions

The term “about” in the present specification means a value within 15%(±15%) of the value recited immediately after the term “about,”including the value equal to the upper limit (i.e., +15%) and the valueequal to the lower limit (i.e., —15%) of this range. For example, thephrase “about 100” encompasses any numeric value that is between 85 and115, including 85 and 115.

The terms “administration” or “administering” compound should beunderstood to mean providing a compound of the present invention to asubject in a form that can be introduced into that subject's body in anamount effective for prophylaxis, treatment, or diagnosis, asapplicable. Such forms may include e.g., oral dosage forms, injectabledosage forms, transdermal dosage forms, inhalation dosage forms, andrectal dosage forms.

An “azapeptide” means a peptide in which one or more α-carbon(s) arereplaced by nitrogen trivalent atom(s).

An “azatide” means a peptide in which all α-carbons are replaced bynitrogen trivalent atoms.

An “aza-amino acid” is defined as an amino acid where the chiralα-carbon atom is replaced by a nitrogen atom.

An “α-nitrogen” means a nitrogen atom bonded to a carbonyl group in anazapeptide or or an azatide. The carbon atom next to the α-nitrogen iscalled the β-carbon.

A “peptidomimetic” means a compound which differs from a peptide that it“mimics” in that one or more α-carbon atoms of the peptide have beenreplaced by a nitrogen atom with or without additional structuralmodification(s) to the side chain(s) of the amino acid residues of thepeptide. The one or more α-carbon atoms of the peptide that are replacedby be, e.g., at the N-termini of the peptide (i.e., the first residue ofthe peptide), at the second residue of the peptide, the C-termini of thepeptide (i.e., the last residue of the peptide), the residue covalentlybound to the C-termini of the peptide, and/or at another residue of thepeptide (e.g., at the site of hydrolysis of the peptide). Despite havinga backbone different from the peptide, the peptidomimetic agentpreserves, extends and/or improves functional activity of the peptide.The peptidomimentic agent is more resistant to degradation than thepeptide and/or has an improved therapeutic activity than the peptideand/or has an improved selectivity for a biological receptor than thepeptide and/or improved affinity to a biological receptor and/orreversed activity at a biological receptor (agonistic activity insteadof antagonist activity or antagonistic activity instead of agonisticactivity).

The term “protected” as it is used herein means that one or moregroup(s) (e.g., —OH) in an amino acid, an aza-amino acid, a peptide, anazapeptide, or a compound is protected with a protecting group (e.g.,Phth, Ddz, etc.). Unless otherwise indicated, the term “protectinggroup” or “protective group,” when used to refer to part of a moleculesubjected to a chemical reaction, means a chemical moiety that is notreactive under the conditions of that chemical reaction, and which maybe removed to provide a moiety that is reactive under those conditions.Protecting groups include, for example, nitrogen protecting groups andhydroxy-protecting groups. Examples of protective group include, e.g.,benzyl, diphenylmethyl, trityl, Cbz, Boc, Fmoc, methoxycarbonyl,ethoxycarbonyl, Phth, Ddz, as well as other protective groups known tothose skilled in the art.

A “side chain radical” of aspartic acid, phenylalanine, alanine,histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, and glutamine havethe following structures:

A“side chain radical of proline” is a secondary amine, in that thealpha-amino group is attached directly to the main chain, making the acarbon a direct substituent of the side chain:

Amino acids which can be used in the present invention are L and D-aminoacids.

The term “pharmaceutically acceptable excipient”, as used herein, means,e.g., a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type, etc. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; cocoa butter and suppositorywaxes; oils such as peanut oil, cottonseed oil, safflower oil, sesameoil, olive oil, corn oil and soybean oil; glycols; such a propyleneglycol; esters such as ethyl oleate and ethyl laurate; agar; bufferingagents such as magnesium hydroxide and aluminum hydroxide; alginic acid;pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol,and phosphate buffer solutions, as well as other non-toxic compatiblelubricants such as sodium lauryl sulfate and magnesium stearate, as wellas coloring agents, releasing agents, coating agents, sweetening,flavoring and perfuming agents, preservatives and antioxidants can alsobe present in the composition, according to the judgment of one skilledin the art of formulations.

Unless otherwise indicated, the terms “prevent,” “preventing” and“prevention” contemplate an action that occurs before a patient beginsto suffer from the symptoms of specified disease or disorder, whichinhibits or reduces the severity of the disease or disorder or of one ormore of its symptoms. The terms encompass prophylaxis.

The compounds of the invention can be used in the form ofpharmaceutically acceptable salts derived from inorganic or organicacids. For clarity, the term “pharmaceutically acceptable salt[s]” asused herein generally refers to salts prepared from pharmaceuticallyacceptable acids or bases including inorganic acids and bases andorganic acids and bases. Suitable pharmaceutically acceptable baseaddition salts include, e.g., metallic salts made from aluminum,calcium, lithium, magnesium, potassium, sodium and zinc or organic saltsmade from lysine, N,N′-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine, meglumine (N-methylglucamine) andprocaine. Suitable non-toxic acids include inorganic and organic acidssuch as acetic, alginic, anthranilic, benzenesulfonic, benzoic,camphorsulfonic, citric, ethenesulfonic, formic, fumaric, furoic,galacturonic, gluconic, glucuronic, glutamic, glycolic, hydrobromic,hydrochloric, isethionic, lactic, maleic, malic, mandelic,methanesulfonic, mucic, nitric, pamoic, pantothenic, phenylacetic,phosphoric, propionic, salicylic, stearic, succinic, sulfanilic,sulfuric, tartaric acid, and p-toluenesulfonic acid. Specific acidsinclude, e.g., hydrochloric, hydrobromic, phosphoric, sulfuric, andmethanesulfonic acids. Examples of specific salts include, e.g.,hydrochloride and mesylate salts. Others are well-known in the art. See,e.g., Remington's Pharmaceutical Sciences, 18^(th) ed. (Mack Publishing,Easton Pa.: 1990) and Remington: The Science and Practice of Pharmacy,19th ed. (Mack Publishing, Easton Pa.: 1995). The preparation and use ofacid addition salts, carboxylate salts, amino acid addition salts, andzwitterion salts of compounds of the present invention may also beconsidered pharmaceutically acceptable if they are, within the scope ofsound medical judgment, suitable for use in contact with the tissues ofhumans and lower animals without undue toxicity, irritation, allergicresponse, and the like, are commensurate with a reasonable benefit/riskratio, and are effective for their intended use. Such salts may alsoinclude various solvates and hydrates of the compound of the presentinvention.

Certain compounds of the present invention may be isotopically labelled,e.g., with various isotopes of carbon, fluorine, or iodine, asapplicable when the compound in question contains at least one suchatom. In preferred embodiments, methods of diagnosis of the presentinvention comprise administration of such an isotopically labelledcompound.

Certain compounds of the present invention may exist as stereoisomerswherein, asymmetric or chiral centers are present. These stereoisomersare “R” or “S” depending on the configuration of substituents around thechiral carbon atom. The terms “R” and “S” used herein are configurationsas defined in IUPAC 1974 Recommendations for Section E, FundamentalStereochemistry, in Pure Appl. Chem., 1976, 45: 13-30. The inventioncontemplates various stereoisomers and mixtures thereof and these arespecifically included within the scope of this invention. Stereoisomersinclude enantiomers and diastereomers, and mixtures of enantiomers ordiastereomers. Individual stereoisomers of compounds of the inventionmay be prepared synthetically from commercially available startingmaterials which contain asymmetric or chiral centers or by preparationof racemic mixtures followed by resolution well known to those ofordinary skill in the art. These methods of resolution are exemplifiedby (1) attachment of a mixture of enantiomers to a chiral auxiliary,separation of the resulting mixture of diastereomers byrecrystallization or chromatography and optional liberation of theoptically pure product from the auxiliary as described in Furniss,Hannaford, Smith, and Tatchell, “Vogel's Textbook of Practical OrganicChemistry”, 5^(th) edition (1989), Longman Scientific & Technical, EssexCM20 2JE, England, or (2) direct separation of the mixture of opticalenantiomers on chiral chromatographic columns or (3) fractionalrecrystallization methods.

Certain compounds of the present invention may exist as cis or transisomers, wherein substituents on a ring may attach in such a manner thatthey are on the same side of the ring (cis) relative to each other, oron opposite sides of the ring relative to each other (trans). Suchmethods are well known to those of ordinary skill in the art, and mayinclude separation of isomers by recrystallization or chromatography. Itshould be understood that the compounds of the invention may possesstautomeric forms, as well as geometric isomers, and that these alsoconstitute an aspect of the invention.

Unless otherwise indicated, a “diagnostically effective amount” of acompound is an amount sufficient to diagnose a disease or condition. Ingeneral, administration of a compound for diagnostic purposes does notcontinue for as long as a therapeutic use of a compound, and could beadministered only once if such is sufficient to produce the diagnosis.

Unless otherwise indicated, a “therapeutically effective amount” of acompound is an amount sufficient to treat a disease or condition, or oneor more symptoms associated with the disease or condition. Theappropriate amount depends upon, among other things, the stage of thedisease or condition; the age of the patient; the weight of the patient;the bioavailability of the compound with respect to a target tissue; theconcentration of compound required in vivo to result in a beneficialeffect relative to control; or the concentration of compound required toresult in a pharmacodynamic effect upon a target amyloid protein at thetarget tissue.

The term “subject” is intended to include living organisms in whichdisease may occur. Examples of subjects generally include mammals, e.g.,humans, monkeys, cows, sheep, goats, dogs, cats, mice, rats, andtransgenic species thereof.

The term “Phth-protected carbamoyl aza-imidazole derivative of anunnatural amino acid” as used herein means an unnatural aza-amino acidcovalently bound (conjugated) to phthalimidyl at its N-terminus and toimidazole at its C-terminus. The unnatural amino acid may be substitutedand unsubstituted.

The term “Phth-protected carbamoyl aza-benzotriazole derivative of anunnatural amino acid” as used herein means means an unnatural aza-aminoacid covalently bound (conjugated) to phthalimidyl at its N-terminus andto benzotriazole at its C-terminus. The unnatural amino acid may besubstituted and unsubstituted.

The term “solid-phase synthesis” means a method in which molecules oratoms (e.g., amino acids, aza-amino acids, etc.) are covalently bound ona solid support material and synthesised step-by-step in a singlereaction vessel utilising selective protecting group chemistry. In thismethod, building blocks are typically protected at all reactivefunctional groups. The order of functional group reactions can becontrolled by the order of deprotection. For example, in an aza-peptidesynthesis, an amino-protected amino acid or an amino-protected aza-aminoacid is bound to a solid phase material (e.g., low cross-linkedpolystyrene beads), forming a covalent bond between the carbonyl groupand the resin, e.g., an amido or an ester bond. Then the amino group isdeprotected and reacted with the carbonyl group of the nextamino-protected amino acid or amino-protected aza-amino acid. This cycleis repeated to form the desired peptide or aza-peptide chain. After allreactions are complete, the synthesised peptide or aza-peptide iscleaved from the bead.

The terms “solution phase synthesis” and “liquid phase synthesis” meansa method in which molecules or atoms (e.g., amino acids, aza-aminoacids, etc.) are synthesized in a solution without being covalentlybound on a solid support material.

The term “synthon” means a building block.

The term “room temperature” means 20° C.

The term “ambient temperature” means 18-28° C.

The terms “parent peptide” and “corresponding peptide” mean a nativepeptide (i.e., natural or convention peptide) that differs from anazapeptide in that one or more of the amino residue(s) of the nativepeptide is (are) replaced by a semicarbazide or a substitutedsemicarbazide (i.e., one or more α-carbon(s) of the native peptide arereplaced by nitrogen trivalent atom(s)) in the azapeptide. Thereplacement may be, e.g., at the N-termini of the peptide (i.e., thefirst residue of the peptide), at the second residue of the peptide, theC-termini of the peptide (i.e., the last residue of the peptide), theresidue covalently bound to the C-termini of the peptide, and/or atanother residue of the peptide (e.g., at the site of hydrolysis of thepeptide).

The term “phthalimidyl” means:

The term “phthaloyl” means:

The abbreviation “N-Phth” means “N-phthalimidyl.”

The abbreviation “Boc” means “tert-butoxycarbonyl.”

The abbreviation “Fmoc” means “9-fluorenylmethoxycarbonyl.”

The abbreviation “Ddz” means“2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl.”

The abbreviation “HOBt” means “1-OH-Benzotriazole.”

The abbreviation “SPPS” means “Solid Phase Peptide Synthesis.”

The abbreviation “TCCA” means “trichloroisocyanuric acid.”

The abbreviation “TBACl” means “tetrabutyl ammonium chloride.”

The abbreviation “Phth” means “phthaloyl.”

The abbreviation “Cbz” means “carboxybenzyl.”

The abbreviation “Pbf” means“2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl.”

The abbreviation “Trt” means “trityl or triphenylmethyl.”

The abbreviation “OtBu” means “O-t-butyl.”

The abbreviation “tBu” means “t-butyl.”

The abbreviation “StBu” means “s-t-butyl.”

The abbreviation “Aloc” means “allyloxycarbonyl.”

The abbreviation “Mtr” means “methoxytrimethylbenzene sulfonyl.”

The abbreviation “Mbh” means “4,4-dimethyloxybenzhydryl.”

The abbreviation “Pmc” means “2,2,5,7,8-pentamethyl-chroman-6-sulfonylchloride.”

The abbreviation “Tmob” means 2,4,6-trimethoxybenzyl.

The abbreviation “OA” means “allyl ester.”

The abbreviation “Acm” means “acetamidomethyl.”

The abbreviation “DEAD” means “Diethyl Azodicarboxylate.”

In peptide chemistry, “deprotection” refers to a process of removing theprotecting groups (e.g., phthaloyl, Boc, Cbz, Fmoc, etc) by a chemicalagent. For example, Boc protecting group could be removed under acidicconditions (e.g., 4M HCl, or neat trifluoroacetic acid TFA); Fmocprotecting group could be removed under basic conditions when pH ishigher than 12 (20% pipyridine/DMF or DCM); and Phthaloyl group can becleaved, e.g., under basic conditions or by the use of hydrazine.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph depicting the stability of bradykinin amide in serumas monitored by area under curve at different timepoints using HPLC atwavelength 220 nm.

FIG. 2 is a graph depicting the higher stability of (1,9)-Aza-Bradykininin serum.

FIG. 3 depicts ¹H NMR spectra of Endomorphin-2 (EM2) andAza-Endomorphin-2.

FIG. 4 contains graphs depicting relative stability of FSSE and K883 inmouse serum as indicated by the area under curve amounts of compoundsmonitored by HPLC at wavelength 210 nm (Y axis).

FIG. 5 is a graph of individual plasma concentrations of K883 afterintravenous administration at 1 mg/kg in male Sprague-Dawley Rats.

FIG. 6 is a graph depicting binding of FSSE and K883 to MD2.

FIG. 7 contains graphs depicting K883 inhibition of HMGB1-induced TNFsecretion.

FIG. 8 contains graphs depicting K883 reduction of APAP-inducedpro-inflammatory cytokines and serum liver enzymes.

FIG. 9 is a graph showing that K883 reduced APAP-induced lethality inmice.

FIG. 10 are graphs depicting binding of K763 and Endomorphin-2 bindingto OPRM-1. K763 binds OPRM-1 with a stronger affinity thanEndomorphin-2.

FIG. 11 is a graph depicting stability of EM2 and K763 in mice.

FIG. 12 is a graph depicting pharmacokinetics (PK) of K763 at 60 min inmice (IP) with acetonitrile extraction.

FIG. 13 is an image of X-ray crystallography of Fmoc-phenylhydrazinecarbazic acid chloride.

FIG. 14A depicts degradation of EM-2, K1167Y and K763 by DPPIV.

FIG. 14B depicts stability of EM-2, K1167Y and K763 in mouse serum.

DETAILED DESCRIPTION

A replacement of one or more α-carbon(s) with nitrogen in a peptideconverts the peptide to an “azapeptide”; and replacement of allα-carbon(s) with nitrogen(s) in a peptide converts the peptide to an“azatide.”

Azapeptides and azatides are peptidomimetics frequently more resistantto enzymatic hydrolysis than the corresponding peptides. The increase inresistance to enzymatic degradation may lead to increased metabolicstability of the compounds and and improved receptor binding. Therefore,azapeptides and azatides are useful tools for drug design, applicationsin medicinal chemistry, and in diagnosis, prevention and treatment ofdiseases.

Azapeptides and azatides may, therefore, be used instead of peptides, aspeptidomimetic agents (“peptidomimetics”). For example, azapeptides andazatides may act used as receptor agonists and antagonists or asprotease inhibitors.

Compounds of Formula (IA), I(B), (II), (III), and (IV) of the presentinvention serve as “building blocks” or synthons for the synthesis ofazapeptides and azatides, including compounds of Formula (V) and (VII),both in solution and solid phase synthesis.

The synthesided azapeptides and azatides, including compounds of Formula(V) and (VII), may, then, be used, e.g., as peptidomimentic agents,e.g., peptidomimetic diagnostic and therapeutic agents, instead ofpeptides in drug discovery and in diagnosis, prevention and treatment ofdiseases.

Compounds of Formula (IA), (IB), (II) and (III)

Compounds of Formula (IA), (IB), (II), and (III) could be used in drugdiscovery, diagnosis, prevention and treatment of diseases, or asbuilding blocks for synthesis of peptidomimetic agents, e.g., for use indrug discovery, diagnosis, prevention and treatment of diseases.

In certain embodiments, a compound of Formula (IA), (IB), (II) and (III)is an imidazole derivative of an aza-amino acid, the imidazolederivative comprising an aza-amino acid covalently bound (conjugated) toa protecting group at its N-terminus and to imidazole at its C-terminus,wherein the aza-amino acid is selected from the group consisting ofaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, and aza-arginine. In some of theseembodiments, the protecting group comprises phthalimide. In additionalembodiments, the protecting group comprises fluorenylmethyloxycarbonyl.In further embodiments, the protecting group comprises 2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl or tert-butoxycarbonyl.

In certain embodiments, the imidazole derivative is stable at 37° C. inan aqueous medium (e.g., an aqueous solution) with a pH of about 7 forat least 30 minutes, 60 minutes, 90 minutes, 1 hour, 2 hours, 3 hours, 4hours or 5 hours. In certain embodiments, the imidazole derivative isstable at 37° C. in mouse serum for at least 30 minutes, 60 minutes, 90minutes, 1 hour, 2 hours, 3 hours, 4 hours or 5 hours.

Compounds of Formula (IA), (IB), (II), and (III) include, e.g.,Phth-protected carbamoyl imidazoles and are Phth-protected carbamoylbenzotriazoles.

Phth-Protected Carbamoyl Imidazoles

In certain embodiments, compounds of Formula (IA), (TB), (II) and (III)are Phth-protected carbamoyl imidazoles. In some of these embodiments,the compounds of Formula (IA), (B), (II) and (III) are selected from thegroup consisting of:

and pharmaceutically acceptable salts thereof, wherein “PG” is H or aprotecting group (e.g., N-phthalimidyl, tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl).

In certain embodiments, compounds of Formula (IA), (IB), (II) and (III)are selected from the group consisting of Phth-aza-asparticacid-carbamoyl imidazole, Phth-aza-phenylalanine-carbamoyl imidazole,Phth-aza-alanine-carbamoyl imidazole, Phth-aza-histidine-carbamoylimidazole, Phth-aza-glutamic acid-carbamoyl imidazole,Phth-aza-tryptophan-carbamoyl imidazole, Phth-aza-valine-carbamoylimidazole, Phth-aza-leucine-carbamoyl imidazole,Phth-aza-lysine-carbamoyl imidazole, Phth-aza-cysteine-carbamoylimidazole, Phth-aza-tyrosine-carbamoyl imidazole,Phth-aza-iso-leucine-carbamoyl imidazole, Phth-aza-arginine-carbamoylimidazole, Phth-aza-glycine-carbamoyl imidazole,Phth-aza-asparagine-carbamoyl imidazole, Phth-aza-glutamine-carbamoylimidazole, and salts thereof.

In certain embodiments, a compounds of Formula (IA), I(B), (II), (III),or (IV) is a Phth-protected carbamoyl imidazole derivative ofaza-proline.

Compounds of Formula (IV)

Compounds of Formula (IV) could be used in drug discovery, diagnosis,prevention and treatment of diseases, or as building blocks forsynthesis of peptidomimetic agents, e.g., for use in drug discovery,diagnosis, prevention and treatment of diseases.

In certain embodiments, a compound of Formula (IV) is a benzotriazolederivative of an aza-amino acid comprising the aza-amino acid covalentlybound (conjugated) to a protecting group at its N-terminus and tobenzotriazole at its C-terminus, wherein the aza-amino acid is selectedfrom the group consisting of aza-glycine, aza-alanine, aza-valine,aza-leucine, aza-isoleucine, aza-proline, aza-phenylalanine,aza-tyrosine, aza-tryptophan, aza-aspartic acid, aza-glutamic acid,aza-aspargine, aza-glutamine, aza-histidine, aza-lysine, andaza-arginine. In certain embodiments, the protecting group comprisesphthalimide. In additional embodiments, the protecting group comprisesfrorenylmethoxycarbonyl (FMOC). In further embodiments, the protectinggroup comprises 2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl.

In certain embodiments, the compound of Formula (IV) is stable at 37° C.in an aqueous medium (e.g., an aqueous solution) with a pH of about 7for at least 30 minutes, 60 minutes, 90 minutes, 1 hour, 2 hours, 3hours, 4 hours or 5 hours.

Phth-Protected Carbamoyl Benzotriazoles

In certain embodiments, the compound of Formula (IV) is a Phth-protectedcarbamoyl benzotriazole.

In certain embodiments, the compound of Formula (IV) is selected fromthe group consisting of:

and pharmaceutically acceptable salts thereof, wherein “PG” is H or aprotecting group (e.g., N-phthalimidyl, tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl).

In certain embodiments, the compound of Formula (IV) is selected fromthe group consisting of Phth-aza-aspartic acid-carbamoyl benzotriazole,Phth-aza-phenylalanine-carbamoyl benzotriazole,Phth-aza-alanine-carbamoyl benzotriazole, Phth-aza-histidine-carbamoylbenzotriazole, Phth-aza-glutamic acid-carbamoyl benzotriazole,Phth-aza-tryptophan-carbamoyl benzotriazole, Phth-aza-valine-carbamoylbenzotriazole, Phth-aza-leucine-carbamoyl benzotriazole,Phth-aza-lysine-carbamoyl benzotriazole, Phth-aza-cysteine-carbamoylbenzotriazole, Phth-aza-tyrosine-carbamoyl benzotriazole,Phth-aza-leucine-carbamoyl benzotriazole, Phth-aza-arginine-carbamoylbenzotriazole, Phth-aza-glycine-carbamoyl benzotriazole,Phth-aza-asparagine-carbamoyl benzotriazole, and salts thereof.

In certain embodiments, a compound of Formula (IA), (IB), (II), (III) or(IV) is a Phth-protected carbamoyl benzotriazole derivative ofaza-proline.

Compounds of Formula (IA), (IB), (II), (III) and (IV) serve as “buildingblock” for compounds of Formula (V) and (VII).

Compounds of Formula (V) and (VII)

Compounds of Formula (V) and (VII) are peptidomimetic analogues ofcompounds of Formula (VI) and and (VIII), respectively. In the preferredembodiments, compounds of Formula (V) and (VII) are more resistant tohydrolysis and/or enzymatic degradation than compounds of Formula (VI)and (VIII).

Compounds of Formula (V) and (VII) may be used to inhibit peptidases,both in vitro and in vivo. The peptidase may, e.g., be an endopeptidase,an exopeptidase, an aspartic protease, a glutamic protease, anasparagine peptide lyase, or a retroviral protease.

In some of these preferred embodiments, compounds of Formula (V) and(VII) are more potent than compounds of Formula (VI) and (VIII), e.g.,due to a better fit into a biological receptor. Compounds of Formula (V)and (VII) could be used, e.g., in drug discovery, diagnosis, preventionand treatment of diseases.

Compounds of Formulas (V) and Formula (VII) may each comprise from 2 to200 carbonyl group(s). For example, compounds of Formula (V) and (VII)may each comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 36,37, 38, 39, 40, 41, 43, 44, 56, or 166 carbonyl groups. In certainembodiments, compounds of Formula (V) and (VII) comprise from 2 to 60carbonyl groups, from 2 to 50 carbonyl groups, from 2 to 40 carbonylgroups, from 2 to 30 carbonyl groups, from 2 to 25 carbonyl groups, from2 to 20 carbonyl groups, from 2 to 15 carbonyl groups, from 2 to 12carbonyl groups, from 2 to 10 carbonyl groups, from 2 to 9 carbonylgroups, from 3 to 40 carbonyl groups, from 3 to 30 carbonyl groups, from3 to 25 carbonyl groups, from 3 to 20 carbonyl groups, from 3 to 15carbonyl groups, from 3 to 12 carbonyl groups, from 3 to 10 carbonylgroups, or from 3 to 9 carbonyl groups.

In certain embodiments, compounds of Formula (V) and (VII) comprise from2 to 200 carbonyl groups and at least one α-nitrogen covalently bound toat least one of said carbonyl groups, and have a greater bioavailability(e.g., oral, transdermal, and/or intranasal) than a peptide structurallydifferent from the compounds of Formula (V) and (VII) only in that thatthe peptide comprises α-carbon instead of said α-nitrogen. In certainembodiments, the α-nitrogen is at the N-termini or C-termini of thecompounds of Formula (V) and (VII). In certain embodiments, theα-nitrogen is one carbonyl group away from the N-termini or C-termini ofthe compounds of Formula (V) and (VII). In certain embodiments, theα-nitrogen is adjacent to the N-termini and the C-termini of thecompounds of Formula (V) and (VII). In certain embodiments, theα-nitrogen is not at the N-termini and not at the C-termini of thecompounds of Formula (V) and (VII) and is more than one carbonyl groupaway for the N-termini and the C-termini. In certain embodiments, theα-nitrogen is at a cleavage or hydrolysis site(s) of the compounds ofFormula (V) and (VII).

In certain embodiments, compounds of Formula (V) and (VII) comprise from2 to 200 carbonyl groups and at least one α-nitrogen covalently bound toat least one of said carbonyl groups, wherein said at least one carbonylgroup is at the N-termini or C-termini residue of the compounds ofFormula (V) and (VII).

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of therapeutic peptides.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of diagnostic peptides.

Compounds of Formula (V) and Formula (VII) may be used in drugdiscovery, diagnosis, prevention and treatment of diseases.

In certain embodiments, compounds of Formula (V) and (VII) comprise abackbone comprising from 2 to 200 carbonyl groups and α-nitrogencovalently bound to at least one of said carbonyl groups, and aretherapeutically effective for the treatment of a disorder in a subject,while a peptide structurally different from the compounds of Formula (V)and (VII) only in that that the peptide comprises α-carbon instead ofsaid α-nitrogen is not therapeutically effective for the treatment ofthe disorder.

In certain embodiments, compounds of Formula (V) and (VII) comprise from2 to 200 carbonyl groups and α-nitrogen covalently bound to at least oneof said carbonyl groups, and have a therapeutic efficacy greater than apeptide structurally different from the compounds of Formula (V) and(VII) only in that the peptide comprises an α-carbon instead of saidα-nitrogen.

In certain embodiments, compounds of Formula (V) and (VII) comprise from2 to 200 carbonyl groups and α-nitrogen covalently bound to at least oneof said carbonyl groups, and have a longer duration of therapeuticactivity than a peptide structurally different from the compounds ofFormula (V) and (VII) only in that that the peptide comprises α-carboninstead of said α-nitrogen.

In certain embodiments, compounds of Formula (V) and (VII) comprise from2 to 75 carbonyl groups and at least one α-nitrogen covalently bound toat least one of said carbonyl groups, and have an in vivo half-lifegreater than a peptide structurally different from the compounds ofFormula (V) and (VII) only in that said at least one α-nitrogen isreplaced with α-carbon.

In certain embodiments, compounds of Formula (V) and (VII) comprise abackbone comprising from 2 to 75 carbonyl groups, wherein at least twocarbonyl groups are covalently bound to a trivalent nitrogen, andcompounds of Formula (V) and (VII) have an in vivo half-life greaterthan a peptide structurally different from the compounds of Formula (V)and (VII) only in that one or more alpha nitrogen(s) of the compounds ofFormula (V) and (VII) is replaced with alpha carbon(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one aza-amino acid, and have an in vivo half-life greater than apeptide structurally different from the compounds of Formula (V) and(VII) only in that the aza-amino acid(s) is replaced with acorresponding amino acid.

In certain embodiments, compounds of Formula (V) and (VII) comprise from2 to 200 carbonyl groups and α-nitrogen covalently bound to at least oneof said carbonyl groups, and are more resistant to protease degradationthan a peptide structurally different from the compounds of Formula (V)and (VII) only in that that the peptide comprises α-carbon instead ofsaid α-nitrogen.

In certain embodiments, compounds of Formula (V) and (VII) comprise from2 to 200 carbonyl groups and α-nitrogen covalently bound to at least oneof said carbonyl groups, and have a greater affinity to a biologicalreceptor than a peptide structurally different from the compounds ofFormula (V) and (VII) only in that that the peptide comprises α-carboninstead of said α-nitrogen.

In certain embodiments, compounds of Formula (V) and (VII) comprisesfrom 2 to 60 carbonyl groups.

In certain embodiments, compounds of Formula (V) and (VII) are linear.

In certain embodiments, compounds of Formula (V) and (VII) are cyclic.

In certain embodiments, compounds of Formula (V) and (VII) arepegylated.

In certain embodiments, compounds of Formula (V) and (VII) areconjugated to an immunoglobulin.

In certain embodiments, compounds of Formula (V) and (VII) compriseα-nitrogen at the N-terminus of its backbone.

In certain embodiments, compounds of Formula (V) and (VII) compriseα-nitrogen at the C-terminus of its backbone

In certain embodiments, compounds of Formula (V) and (VII) comprise twocarbonyl groups and two α-nitrogens.

In certain embodiments, compounds of Formula (V) and Formula (VII)comprise three carbonyl groups and one α-nitrogen.

In certain embodiments, a compound of Formula (V) or a compound ofFormula (VII) comprises three carbonyl groups and two α-nitrogens.

In certain embodiments, compounds of Formula (V) and (VII) comprisethree carbonyl groups and three α-nitrogens.

In certain embodiments, compounds of Formula (V) comprise four carbonylgroups and one α-nitrogen.

In certain embodiments, compounds of Formula (V) comprise four carbonylgroups and two α-nitrogens.

In certain embodiments, compounds of Formula (V) and (VII) comprise fourcarbonyl groups and three α-nitrogens.

In certain embodiments, compounds of Formula (V) and (VII) comprise fourcarbonyl groups and four α-nitrogens.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a 2 to 200 amino acid peptide comprising an amino acidselected from the group consisting of glycine, alanine, valine, leucine,isoleucine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid,glutamic acid, aspargine, glutamine, histidine, lysine, and arginine;the aza-analogues differing from the amino acid peptide in that that theamino acid of the peptide is replaced with a corresponding aza-aminoacid.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a 2 to 200 amino acid peptide comprising an amino acidselected from the group consisting of glycine, alanine, valine, leucine,isoleucine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid,glutamic acid, aspargine, glutamine, histidine, lysine, and arginine,wherein the analogue includes at least one corresponding aza-amino acidof the amino acid.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a 2 to 200 amino acid peptide, the 2 to 200 amino acidpeptide comprising amino acids selected from the group consisting ofglycine, alanine, valine, leucine, isoleucine, proline, phenylalanine,tyrosine, tryptophan, aspartic acid, glutamic acid, aspargine,glutamine, histidine, lysine, arginine, the analogue differing from theamino acid peptide in that that the aza-analogues comprise an aza-aminoacid instead of at least one of the amino acids, wherein theaza-analogues comprise aza-glycine instead of glycine, and/or theaza-analogues comprise aza-alanine instead of alanine, and/or theaza-analogues comprise aza-valine instead of valine, and/or theaza-analogues comprise aza-leucine instead of leucine, or/and theaza-analogues comprise aza-isoleucine instead of iso-leucine, and/or theaza-analogues comprise aza-proline instead of proline, and/or theaza-analogues comprise aza-phenylalanine instead of phenylalanine,or/and the aza-analogues comprise comprises aza-tyrosine instead oftyrosine, and/or the aza-analogues comprise aza-tryptophan instead oftryptophan, or/and the aza-analogues comprise aza-aspartic acid insteadof aspartic acid, and/or the aza-analogues comprise aza-glutamic acidinstead of glutamic acid, and/or the aza-analogues compriseaza-aspargine instead of aspargine, and/or the aza-analogues compriseaza-glutamine instead of glutamine, and/or the aza-analogues compriseaza-histidine instead of histadine, and/or the aza-analogues compriseaza-lysine instead of lysine, and/or the aza-analogues compriseaza-arginine instead of arginine.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide comprising from 2 to 50 amino acids selectedfrom the group consisting of glycine, alanine, valine, leucine,isoleucine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid,glutamic acid, aspargine, glutamine, histidine, lysine, arginine, and atleast 2, 3, 4, 5, 6, 7, 8, 9 or 10 of the amino acids are replaced withcorresponding aza-amino acids. In some of these embodiments, thereplaced amino acid is the first amino acid of the peptide. In some ofthese embodiments, the replaced amino acid is the second amino acid ofthe peptide. In some of these embodiments, the replaced amino acid isthe last amino acid of the peptide. In some of these embodiments, thefirst and the last amino acids of the peptide are both replaced withcorresponding aza-amino acids. In some of these embodiments, the aminoacids of the peptide adjacent to the N-termini and the C-termini of thepeptide are both replaced with corresponding aza-amino acids.

In certain embodiments, the last amino acid of the peptide is selectedfrom the group consisting of aspartic acid, phenylalanine, and arginine.

In certain embodiment, the first amino acid of the peptide is selectedfrom the group consisting of tyrosine, phenylalanine, and arginine.

In certain embodiments, the first and the last amino acid of the peptideare the same.

In certain embodiments, the first and the last amino acids of thepeptide are different.

In certain embodiments, compounds of Formula (V) and (VII) are notazatides.

In certain embodiments, compounds of Formula (V) and (VII) comprise anamino acid selected from the group consisting of cysteine, methionine,serine and threonine.

In certain embodiments, compounds of Formula (V) and (VII) comprises atleast one, at least two or at least three aza-glycine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-alanine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-valine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-leucine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-isoleucine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-proline(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-phenylalanine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-tyrosine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-tryptophan(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-aspartic acid(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-glutamic acid(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-aspargine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-glutamine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-histidine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-lysine(s).

In certain embodiments, compounds of Formula (V) and (VII) comprise atleast one, at least two or at least three aza-arginine(s).

In certain embodiments, compounds of Formula (V) and (VII) compriseaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, or aza-arginine on their N-termini and/orC-termini.

In certain embodiments, compounds of Formula (V) and (VII) compriseaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, or aza-arginine on their N-termini and/orC-termini, and are aza-analogues of a therapeutic peptide, and have agreater bioavailability (e.g., oral, transdermal, and/or intranasal)than the therapeutic peptide (in its unaltered state).

In certain embodiments, compounds of Formula (V) and (VII) compriseaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, or aza-arginine on their N-termini and/orC-termini, and are aza-analogues of a therapeutic peptide, maintain thetherapeutic efficacy of the therapeutic peptide and have an in vivohalf-life greater than the in vivo half-life of the therapeutic peptide.In some of these embodiments, the in vivo half-life of the compounds ofFormula (V) and (VII) is greater than twice of the in vivo half-life ofthe therapeutic peptide. In some of these embodiments, the in vivohalf-life of the compounds of Formula (V) and (VII) is three timesgreater than the in vivo half-life of the therapeutic peptide. Inadditional embodiments, the in vivo half-life of the compounds ofFormula (V) and (VII) is four times greater than the in vivo half-lifeof the therapeutic peptide. In yet additional embodiments, the in vivohalf-life of the compounds of Formula (V) and (VII) is five timesgreater than the in vivo half-life of the therapeutic peptide. Infurther embodiments, the in vivo half-life of the compounds of Formula(V) and (VII) is six times greater than the in vivo half-life of thetherapeutic peptide. In additional embodiments, the in vivo half-life ofthe compounds of Formula (V) and (VII) is ten times greater than the invivo half-life of the therapeutic peptide. The in vivo half-life of thecompounds of Formula (V) and (VII) may, e.g., be from about 1 minute toabout 72 hours.

In certain embodiments, compounds of Formula (V) and (VII) compriseaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, or aza-arginine on their N-termini and/orC-termini, are aza-analogues of a therapeutic peptide and have a longerduration of therapeutic activity than the therapeutic peptide.

In certain embodiments, compounds of Formula (V) and (VII) compriseaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, or aza-arginine on their N-termini and/orC-termini, are aza-analogues of a therapeutic peptide and are moreresistant to protease degradation than the therapeutic peptide.

In certain embodiments, compounds of Formula (V) and (VII) compriseaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, or aza-arginine on their N-termini and/orC-termini, are aza-analogues of a therapeutic peptide and have a greateraffinity to a biological receptor than the therapeutic peptide.

In certain embodiments, compounds of Formula (V) and (VII) are compoundsof formula:

wherein Z₁ and Z_(n) is each independently C or N;

R is hydrogen, —NH₂, —NNH₂, —CONH₂, —COOR₃, —COOH, —COH, —COC₁-C₄ alkyl,—COC₁-C₄ haloalkyl, —OH, an amino acid, an aza amino acid, a 2 to 60-merpeptide, a 2 to 60-mer aza peptide, a 2 to 60-mer azatide;

R₁ and R₂ is each independently selected from the group consisting of H,methyl, isopropyl, isobutyl, benzyl, and side chain radicals of asparticacid, phenylalanine, alanine, histidine, glutamic acid, tryptophan,valine, leucine, lysine, methionine, threonine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, serine, and glutamine;

Q is NH₂ or OH;

at least one of Z₁ and Z_(n) is N; and

n is an integer from 1 to 200.

For example, n could be 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65,66, 67, 68, 69, 70, etc. In certain embodiments, n is 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. The side chainradicals may be unsubstituted or substituted with one or more of thefollowing: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl),hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl(e.g., a chloromethyl, a fluromethyl, etc.).

Di-Mer Azapeptides

In certain embodiments, a compound of Formula (V) or (VII) is a compoundof formula:

wherein Z₁ and Z₂ is each independently C or N;

R₁ and R₂ is each independently selected from the group consisting of H,methyl, isopropyl, isobutyl, benzyl, and side chain radicals of asparticacid, phenylalanine, alanine, histidine, glutamic acid, tryptophan,valine, leucine, lysine, methionine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, serine, and glutamine;

Z is NH₂ or OH; and

at least one of Z₁ and Z₂ is N. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.). In certain embodiments, R₁ and R₂ is eachindependently selected from the group consisting of H, H₂ and side chainradicals of aspartic acid, histidine, glutamic acid, tryptophan, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, asparagine, and glutamine.

In certain embodiments, a compound of Formula (V) or (VII) is adi-azatide of a compound of Formula (IX)

or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom the group consisting of unsubstituted and substituted side chainradicals of aspartic acid, phenylalanine, alanine, histidine, glutamicacid, tryptophan, valine, leucine, lysine, methionine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, and glutamine.

The di-azatides may, e.g., be prepared by a solution phase or a solidphase synthesis:

The di-azatides may be prepared both with C-to-N terminal constructionand N-to-C terminal construction.

C-to-N-Terminal Construction

In the method of C-to-N-terminal construction, the di-azatide amide canbe made by coupling of hydrazine amide (1) with acid chloride (2) inDCM/toluene at about 25° C. or 50° C. to yield the N-Fmoc protectedDi-azatide amide (3) which may be de-protected, e.g., with piperidine toyield the final Di-azatide amide (4):

The following General Procedure may be used for coupling of hydrazineamide with acid chloride (C-to-N-terminal construction): The solution ofacid chloride (0.367 mmol) and hydrazine amide (0.367 mmol) in anhydrousDCM (3 mL) and anhydrous toluene (3 mL) is stirred at about 50° C. undernitrogen for, e.g., about 15 hours. The mixture is concentrated todryness and then the crude product is purified by flash silica gelcolumn chromatography eluting with hexane/EtOAc mixtures to afford theproducts as a white solids or clear oils in, e.g., 50-70% yield.

N-to-C-Terminal Construction

In the method of N-to-C-terminal construction to the Di-azatide (4) byformation of protected carbazide (5) from acid chloride (2) and thencoupled with an appropriate aldehyde to form an acyl hydrazone which maybe reduced by, e.g, the catalytic hydrogenation and hydride addition toyield Di-azatide (6). Chlorosulfonyl isocyanate (CSI) may be used toconvert amine into the corresponding amide which may then de-protected,e.g., with piperidine to yield the final Di-azatide amide (4):

wherein R is selected from the group consisting of unsubstituted andsubstituted side chain radicals of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, and glutamine.

The following General Procedure may be used for coupling of protectedcarbazide with aldehydes (N-to-C-terminal construction): To a solutionof protected carbazide (0.46 mmol) and aldehyde (0.69 mmol) in anhydrousmethanol (20 mL) triethyl amine (60 uL until pH=7) is added and followedby anhydrous MgSO₄ (200 mg). The mixture is stirred at about 55° C.under nitrogen for, e.g., about 1 hour. Then, NaCNBH₃ (2.3 mmol) isadded, followed by acetic acid (2.3 mmol). The mixture is stirred at 80°C. under nitrogen for, e.g., about 15 hours then concentrated to drynessand partitioned between water (100 mL) and EtOAc (100 mL). The aqueouslayer is extracted with EtOAc (2×50 mL) and the combined organic phaseare washed with brine (50 mL), dried over Na₂SO₄, filtered andconcentrated to afford the crude product which is purified by flashsilica gel column chromatography eluting with hexane/EtOAc mixtures toafford the products as white solids or clear oils, e.g., in 40 to 60%yield.

The following General Procedure may also be used for coupling ofhydrazine with acid chloride: To a solution of acid chloride (0.367mmol) and hydrazine (3.67 mmol) in anhydrous DCM (6 mL) was addedN,N,N′, N′-Tetramethyl-1,8-naphthalenediamine (0.734 mmol). The solutionwas stirred at about 25° C. under nitrogen for, e.g., about 15 hours,concentrated to dryness and partitioned between 0.5N HCl (20 mL) andEtOAc (20 mL). The aqueous layer is extracted with EtOAc (2×25 mL) andthe combined organic phase are washed with brine (25 mL), dried overNa₂SO₄, filtered and concentrated to afford the crude product which waspurified by flash silica gel column chromatography eluting withhexane/EtOAc mixtures to afford the products as a white solids or clearoils, e.g., in 70-80% yield.

The following General Procedure may be used for removing Fmoc group withpiperidine: The solution of N-Fmoc protected azatide (1.0 mmol) inpiperidine (5 mL) is stirred at about 35° C. under nitrogen for about 15mins. The mixture is concentrated to dryness and then the crude productis purified by flash silica gel column chromatography eluting withhexane/EtOAc mixtures to afford the products as a white solids, e.g., in90-95% yield.

Compounds of Formula (IX) can be used as diagnostic peptidomimeticagents, therapeutic peptidomimetic agents, and in drug discovery, e.g.,to synthese longer peptidomimetic agents (e.g., tri-azatides,tetra-azatides, etc.).

Tri-Mer Azapeptides and Azatides

In certain embodiments, a compound of Formula (V) or (VII) is a compoundof:

wherein Z₁, Z₂ and Z₃ is each independently C or N;

R₁, R₂, and R₃ is each independently selected from the group consistingof H, methyl, isopropyl, isobutyl, benzyl, and side chain radicals ofaspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, threonine tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, serine, and glutamine;

Q is NH2 or OH; and

at least one of Z₁, Z₂ and Z₃ is N. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.). In certain embodiments, R₁, R₂, andR₃ is each independently selected from the group consisting of H, H₂ andside chain radicals of aspartic acid, histidine, glutamic acid,tryptophan, lysine, methionine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, asparagine, andglutamine.

In certain embodiments, a compound of Formula (V) or (VII) is atri-azatide of Formula (X):

or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom the group consisting of unsubstituted and substituted side chainradicals of aspartic acid, phenylalanine, alanine, histidine, glutamicacid, tryptophan, valine, leucine, lysine, methionine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, proline, and glutamine.

The tri-azatides may be prepared, e.g., by coupling of di-azatide amide(7) with acid chloride (2) in DCM/toluene at about 50° C. to yield theN-Fmoc protected Tri-azatide amide (8):

wherein R is selected from the group consisting of unsubstituted andsubstituted side chain radicals of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, and glutamine.

The following General Procedure may be used to remove Fmoc group withpiperidine: The solution of N-Fmoc protected azatide (1.0 mmol) inpiperidine (5 mL) is stirred at about 35° C. under nitrogen for about 15mins. The mixture is concentrated to dryness and then the crude productis purified by flash silica gel column chromatography eluting withhexane/EtOAc mixtures to afford the products as white solids, e.g., in90-95% yield.

Compounds of Formula (X) can be used as diagnostic peptidomimeticagents, therapeutic peptidomimetic agents, and in drug discovery, e.g.,to synthesize longer peptidomimetic agents (e.g., tetra-azatides, etc.).

Four-Mer Azapeptides

In certain embodiments, a compound of Formula (V) or (VII) is a compoundof formula:

wherein Z₁, Z₂, Z₃, and Z₄ is each independently C or N;

R₁, R₂, R₃, and R₄ is each independently selected from the groupconsisting of H, methyl, isopropyl, isobutyl, benzyl, and side chainradicals of aspartic acid, phenylalanine, alanine, histidine, glutamicacid, tryptophan, valine, leucine, lysine, methionine, tyrosine,isoleucine (including, R-isoleucine, S-isoleucine and RS-isoleucine),arginine, glycine, asparagine, serine, and glutamine;

Q is NH2 or OH; and

at least one of Z₁, Z₂, Z₃, and Z₄ is N. The side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.). In certain embodiments, R₁, R₂, R₃,and R₄ is each independently selected from the group consisting of H, H₂and side chain radicals of aspartic acid, histidine, glutamic acid,tryptophan, lysine, methionine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, asparagine, andglutamine.

In certain embodiments, a compound of Formula (V) or (VII) is a compoundof formula:

or a pharmaceutically acceptable salt thereof.

In certain embodiments, a compound of Formula (V) or (VII) is a compoundof formula:

or a pharmaceutically acceptable salt thereof.

Nine-Mer Azapeptides

In certain embodiments, a compound of Formula (V) or (VII) is a compoundof formula:

wherein Z₁, Z₂, Z₃, Z₄, an Zs is each independently or N;

Q is NH₂ or OH;

R₁, R₂, R₃, and R₄ is each independently selected from the groupconsisting of H and side chain radicals of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, serine, andglutamine. The side chain radicals may be unsubstituted or substitutedwith one or more of the following: a halogen (Cl, F, or Br), a C₁-C₆alkyl (e.g., methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl,propoxyl, a C₁-C₆ haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.)or a protecting group (e.g., Phth, Boc, Fmoc, Ddz, etc.). In certainembodiments, R₁, R₂, R₃, and R₄ is each independently selected from thegroup consisting of H, methyl, isopropyl, isobutyl, benzyl, and sidechain radicals of aspartic acid, histidine, glutamic acid, tryptophan,lysine, methionine, tyrosine, isoleucine (including, R-isoleucine,S-isoleucine and RS-isoleucine), arginine, asparagine, and glutamine;and

at least one of Z₁, Z₂, Z₃, Z₄, and Zs is N.

In certain embodiments, a compound of Formula (V) or (VII) is a compoundof formula:

or a pharmaceutically acceptable salt thereof.

Utility of Compounds of Formula (V) and (VII)

In certain embodiments, compounds of Formula (V) and (VII) are used indrug discovery, e.g., to provide a library of compounds suitable for thediagnosis, prevention or treatment of a pathological condition(s).

In certain embodiments, compounds of Formula (V) and (VII) are used asdiagnostic agents.

In certain embodiments, compounds of Formula (V) and (VII) are used astherapeutic agents.

In certain embodiments, compounds of Formula (V) and (VII) are used asinhibitors of proteases (e.g., an endopeptidase, an exopeptidase, anaspartic protease, a glutamic protease, an asparagine peptide lyase, aretroviral protease, etc.).

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for the treatment of acne, acromegaly,alopecia, anemia, asthma, cancer, age-related macular degeneration, bonecysts, dental caries, cognitive enhancement, cystic fibrosis,chemoprevention, Cushing's syndrome, anorexia nervosa, depression,obsessive-compulsive disorder, diabetic retinopathy, diabetic macularedema, diabetic nephropathy, dyspepsia, brain edema, epilepsy, renalfailure, gingivitis, lupus erythematosus, chronic lyphocytic leukemia,obesity, estrogen deficiency, emesis, endometriosis, endometrialthinning, gastrointestinal disorders, gigantism, bone injuries, toothrestoration, heart failure, myocardial infarction, cerebrovascularischemia, ischemia, unstable angina pectoris, hypertension, isolatedsystolic hypertension, cardiovascular disease, coronary disorder,atherosclerosis, peripheral artery disease, arrhythmia, pain,vasodilatory hypotension, intradialytic hypotension, stroke, sepsis,thromboembolism, restenosis, hypercalcemia, inflammation, type 1diabetes, type 2 diabetes, wound healing, eryrthropietic protoporphyria,photodamage, actinic keratosis, myasthenia gravis, multiple sclerosis,transplant rejection, lipid metabolism disorder, malnutrition, HIV,hepatitis, herpes, glaucoma, osteoporosis, erectile dysfunction,rheumatoid arthritis, Alzheimer's disease, dermal scarring, kelidscarring, atopic dermatitis, impetigo, uveitis, uterine contractions,acute coronary syndrome, thrombosis, neutropenia, thrombocytopenia(e.g., heparin-induced thrombocytopenia), female sexual dysfunction,female infertility, postpartum uterine atony, postpartum hemorrhagebleeding, Paget's disease, gastric disorders, Gram negative bacterialinfection, mycosesm, bacteremia, candidemia, diarrhea, candida ablicantsinfection, vulvovaginal candidiasis, pancreatic dysfunction, benignprostatic hyperplasia, uterine fibroids, growth disorder, metabolicsyndrome, metabolic disorder, HIV-associated lipodystrophy, cachexia,Factor VIII deficiency, multiple sclerosis, Graft versus host disease,epilepsy, Parkinson's disease, schizophrenia, functional bowel disease,inflammatory bowel disease, irritable bowel syndrome, ulcerativecolitis, Crohn's disease, Celiac disease, short bowel syndrome, ileus,systemic inflammatory response syndrome, brain edema, head injury,precocious puberty, polycystic ovary syndrome, uterine fibroids,nocturia, diabetis insipidus, enuresis, polyuria, primary nocturnalenuresis, Von Willebrand's disease, Hemophilia, hemopoietic disorder,female contraception, male contraception, scleroderma, diabetic footulcer, septic shock, cognition disorder, dementia, HIV-associateddementia, mild cognitive impairment, systemic lupus erythematosus,somatotropin deficiency, muscle wasting, skin disorders, reperfusioninjury, inhibition of premature LH surges, Leukopenia, drug inducedfungal infection, onychomycosis, immune disorder, viral infection,immune deficiency, Huntington's chorea, motor neuron disease,neurodegenerative disorder, psoriasis, tuberculosis, respiratory tractdisorders, postoperative infections, lung disorders, radiation sickness,transplant rejection, hereditary angioedema, rhinitis, allergy, asthma,osteoarthritis, liver cirrhosis, respiratory distress syndrome,stomatitis, pneumonia, nutritional disorders, short stature, respiratorydistress syndrome, lung malformation, postoperative ileus, vasoactiveintestinal peptide, stem cell mobilisation, stem cell transplantation,myelofibrosis, catheter infection, rosacea, otitis, conjunctivitis,neuropathy, control of bleeding, delivery induction, labor initiation,labor stimulation, pemphigus vulgaris, muscle weakness, immunethrombocytopenic purpura, myelodysplastic syndrome, spinal fusion,chronic wounds, bleeding esophageal varices, spinocerebellardegeneration, renal disease, hepatorenal syndrome, insomnia, influenzavirus, aspergillus infection, lung infection, primaryimmunodeficiencies, angiogenesis disorder, recurrent autoimmunecytopenia, decubitus ulcer, varicose ulcer, epidermolysis bullosa, eyesurgery, deafness, or labyrinthitis (inflammation of inner ear).

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of coronary thrombosis.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of Clostridiumdefficile-associated diarrhea.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of neonatal respiratory distresssyndrome.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of a ventricular arrhythmia.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of atrial fibrillation.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of respiratory disorder.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of adult neonatal distresssyndrome.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of allergic rhinitis.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of allergic conjunctivitis.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of postmenoposal osteoporosis.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of chemotherapy induceddiarrhea.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of a bone fracture.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for treatment of Staphylococcus aureusinfection.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for the treatment of breast cancer, colorectalcancer, carcinoid cancers, carcinoma, renal cell carcinoma, endometrialcarcinoma, glioma, glioblastoma, hepatocellular carcinoma, lymphoma,non-small lung cancer, ovarian cancer, gastrointestinal cancer,pancreatic cancer, prostate cancer, sarcoma, solid tumors, metastaticmelanoma, multiple myeloma, malignant melanoma, neuroblastoma, skincancer, non-hodgkin lymphoma, small-cell lung cancer, non-small-lungcancer, mesothelioma, pancreatic cancer, hematological neoplasm,neuroendocrine tumors, pituitary cancer, uterine cancer, orosteosarcoma.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for the treatment of neuropathic pain,neuralgia, postoperative pain, cancer pain, inflammatory pain, or painwithout inflammation.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for the treatment of diabetic neuropathy.

In certain embodiments, compounds of Formula (V) and (VII) aretherapeutically effective for the treatment of hypoplastic anemia.

In certain embodiments, a compound of Formula (V) or a compound ofFormula (VII) compounds of Formula (V) and (VII) are for the treatmentof hepatitis A, hepatitis B, or hepatitis C.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of antagonists of ANP receptor, AT receptor,B-cell activating factor, B2 receptor, BB2 receptor, N-Cadherin, calciumchannel, CCRP5 receptor, CALC receptor and RAMPs, CD4 receptor, C5areceptor, CD29 receptor, CXCR4 receptor, GCG receptor, Erb-3 receptor,GnRH R, GP IIb IIIa receptor, integrin alpha-5/beta-3, integrinalpha-4/beta-1, NMDA receptor, Nicotinic ACH receptor, OT receptor, PTHreceptor, SST receptor, TAC1 receptor, TAC2 receptor, TBXA2 receptor,VEGF receptor, VE-Cadherin receptor, and zonulin receptor.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of agonists of an ANP receptor, AVP receptor,B2 receptor, BNP receptor, CCK receptor, CALC receptor, CALC receptorand RAMPs, CRH receptor, CD 36 receptor, CD110 receptor, CXCR4 receptor,EPO receptor, FGF receptor, ET-B receptor, GCG receptor, GH receptor,GNRH receptor, GnRH R receptor, GPL-1 receptor, GPL-2 receptor, GHSreceptor, GPR54, Guanylate cyclase-C, IL2 receptor, IGF-1 receptor, PGE2receptor, NGF receptor, NMDA receptor, NOD protein receptor, NPYreceptor, MC receptor, M1 receptor, NTS1 receptor, NK receptor, PTHreceptor, Delta opioid receptor, Kappa opioid receptor, Mu opioidreceptor, ORL1 receptor, OGF receptor, OT receptor, PAR receptor, SCTreceptor, SST receptor, SST receptor and Dopamine D2 receptor, TRHreceptor, VPAC receptor, and RAMPs receptor.

In certain embodiments, compounds of Formula (V) and (VII) areimmunomodulators.

In certain embodiments, compounds of Formula (V) and (VII) modulate NODprotein.

In certain embodiments, compounds of Formula (V) and (VII) modulate STATprotein.

In certain embodiments, compounds of Formula (V) and (VII) modulateactin.

In certain embodiments, compounds of Formula (V) and (VII) modulate PTHreceptor.

In certain embodiments, compounds of Formula (V) and (VII) modulate GHSreceptor.

In certain embodiments, compounds of Formula (V) and (VII) modulatetubulin.

In certain embodiments, compounds of Formula (V) and (VII) inhibit aprotease.

In certain embodiments, compounds of Formula (V) and (VII) are JNKinhibitors.

In certain embodiments, compounds of Formula (V) and (VII) inhibit HIVTat protein.

In certain embodiments, compounds of Formula (V) and (VII) are thrombininhibitors.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of HDAC.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of protein kinase C delta.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of enkephalinase.

In certain embodiments, compounds of Formula (V) and (VII) stimulateERK.

In certain embodiments, compounds of Formula (V) and (VII) activateprotein kinase C epsilon.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of renin.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsfor DNA.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsfor GP41.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsfor angiopoietin 2.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsfor CD4.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsin cholesterol transport.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsof TGF beta 1.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsof HIV p24.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsof iron.

In certain embodiments, compounds of Formula (V) and (VII) areinsulinotropic.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsof Aβ42.

In certain embodiments, compounds of Formula (V) and (VII) are ligandsof glycosphingolipids.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of Serine-Threonine kinase.

In certain embodiments, compounds of Formula (V) and (VII) are chloridechannel inhibitors.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of compliment C3 enzyme.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of beta-secretase.

In certain embodiments, compounds of Formula (V) and (VII) areinhibitors of matrix metalloproteinase-9.

In certain embodiments, compounds of Formula (V) and (VII) aremodulators of a gap junction.

In certain embodiments, compounds of Formula (V) and (VII) are used asadjuvants to a local anesthetic.

In certain embodiments, a compound of Formula (V) is desmopressin.

In certain embodiments, compounds of Formula (V) and (VII) areconjugated to a small molecule.

In certain embodiments, compounds of Formula (V) and (VII) are dopamineagonists.

In certain embodiments, compounds of Formula (V) and (VII) are dopamineantagonists.

In certain embodiments, compounds of Formula (V) and (VII) areconjugated to a phospholipid.

In certain embodiments, compounds of Formula (V) and (VII) aresurfactants.

In certain embodiments, compounds of Formula (V) and (VII) are GnRHreceptor targeting ligands.

In certain embodiments, compounds of Formula (V) and (VII) exclude[azaVal3]-angiotensin II; azaAsn5-oxytocin; azaGly9-oxytocin;[azaAsn5]-eledoisin; azaGly10-analogues of leutinizizinghormone-releasing hormone (LH-RH) (azaGy10-LH-RH, [D-Ser(But)6,azaGly10]-LH-RH); azaGly analoges of encephalin; aza analogues of thenative peptide ligand, Arg-Gly-Asp (RGD) (i.e., azaAla-RGD; azaGly-RGD,and azaGly aminopyridine analog of RGD); azaPhe4 analog of the growthhormone releasing peptides (GIRP-6) [His-D-Trp-Ala-Trp-D-Phe-Lys-NH2];azaD-Phe and azaArg analogues of melanocortin receptor (MCR)[Ac-His-D-Phe-Arg-Trp-NH2]; analogue of MCR in which Trp was replacedwith aza-Nal-1, aza-Nal-2 and aza-Bip; azaGly33 analog of calcitoningene-related peptide (CGRP); azaTyr analog of the insulin receptortyrosine kinase (IRTK); Boc-azaPhe-trans-Chx-Arg-CONH(s-PhEt);Marocyclic azapeptide inhibitor, BILN 2061, and Atazanavir (CGP 7354 orBMS-232332) (Reyataz®), and azapeptides inhibitors of serine andcysteine proteases described in [27]-[35], all herein incorporated byreference.

The concept of employing an electrophilic aza-residue at the P1 positionwas effective against cysteine proteases such as potent inhibitors ofpapain, cathepsin B, calpains. Caspase-1 and the 3C protease from humanrhinovirus strain 1, without having inhibitory activity on the serinepeptidases, trypsin and porcine pancreatic elastase. (Reference 28).

In certain embodiments, compounds of Formula (V) and (VII) exclude thefollowing compounds:

In certain embodiments, compounds of Formula (V) and (VI) are selectedfrom the group consisting of aza-analogues of A-6, A-623 (AMG-623),A-71378, A-75998, Abarelix (PPI-149), ABT-510, AC-100, AC-162352 (PYY3-36), AC-253, AC-2592, AC-625, ACV-1, ADH-1, AEZS-108 (AN-152)(ZEN-008), AF-37702, Afamelanotide (EP-1647) (CUV-1647) (Melanotan I),AG2/102, AG-284, AI-502, AKL-0707 (LAB GHRH), Albiglutide (GSK-716155),Albuvirtide, ALG-889, Alloferon, Allotrap 2702 (B-2702), ALTY-0601,ALX-40-4C, Ambamustine (PTT-119), Anaritide, Antagonist G (PTL-68001),AOD-9604, APL-180, ATN-161, Atosiban (ORF-22164), Atriopeptin, Aviptadil(PSD-510), Avorelin (EP-23904), AZD-2315, Azetirelin (YM-14673),AZX-100, B27PD, BA-058, Barusiban (FE-200400), BAY-73-7977, BDM-E,BGC-728, BIM-23190, BIM-44002, BIO-1211, Bivalirudin (BG-8865),BMS-686117, Bremelanotide (PT-141), BRX-0585, Buserelin, Calcitonin(Human), Calcitonin (Salmon), Carbetocin, Carfilzomib (PR-171),Cargutocin (Y-5350), Carperitide (SUN-4936), Casokefamide, CB-182804,CB-183315, CBP-501, CBT-101, CCK (25-33), CD-NP, Cemadotin (LU-103793),Cetrorelix (NS-75), CG-77X56, CGRP (LAB-CGRP), Chlorotoxin (TM-601),Cilengitide (EMD-121974) (EMD-85189), CJC-1008 (DAC: Dynorphin A),CJC-1131 (DAC:GLP-1), CJC-1134 (PC-DAC) (Exendin-4), CJC-1295 (DAC:GRF),Cnsnqic-Cyclic (802-2), Compstatin (POT-4), Conantokin G, Contulakin G(CGX-1007), Corticorelin (NEU-3002), CP-95253, C-peptide (SPM-933),CR-665, CR-845, CTCE-0214, CTCE-9908, CTS-21166 (ASP-1702) (ATG-Z1)(OM-00-3) (OM-99-2), CVX-045, CVX-060, CVX-096 (PF-4856883), CZEN-002,D-4F (APP-018), Danegaptide (ZP-1609) (WAY-261134) (GAP-134),Davalintide (AC-2307), Davunetide (AL-108) (AL-208), Degarelix (FE200486), Delmitide (RDP-58), Deltibant (CP-0127), Deslorelin,Desmopressin, Detirelix (RS-68439), DG-3173 (PTR-3173), Didemnin B(NSC-325319), Dirucotide (MBP-8298) Disitertide (NAFB-001) (P-144),DMP-728 (DU-728), dnaJP1 (AT-001), Dopastatin (BIM-23A760), DPK-060,DRF-7295, DSC-127, Dynorphin A, E-2078, EA-230, Ebiratide (Hoe-427),Edotreotide (SMT-487), Edratide (TV-4710), Efegatran (LY-294468),Elcatonin, Eledoisin (ELD-950), Elisidepsin (PM-02734), EMD-73495,Enfuvirtide (T-20), EP-100, EP-51216 (EP-51389), Eptifibatide (C₆₈-22),ET-642 (RLT-peptide), ETRX 101, Examorelin (EP-23905) (MF-6003),Exenatide (AC-2993) (LY-2148568), Exsulin (INGAP Peptide), F-991,FAR-404, FE 202158, Felypressin, FGLL, Frakefamide (LEF-576) (SPD-759)(BCH-3963), FX-06, Ganirelix (Org-37462) (RS-26306), Glaspimod(SKF-107647), Glatiramer (COP-1), Glucagon, Glucosamyl muramyltripeptide, Glutoxim (NOV-002), Glypromate, GMDP, Golotimod (SCV-07),Goralatide (BIM-32001), Goserelin (ICI-118630), GPG-NH2, GTP-200,GTP-300, H-142, Hemoparatide (PTH(1-37)), Hexapeptide copper II(PC-1358), Histrelin, hLF(1-11), HP-228, I-040302 (KUR-112), Icatibant(JE-049) (HOE-140), lcrocaptide (ITF-1697), IMX-942, lpamorelin(NNC-26-0161), IPP-201101, Iseganan (IB-367), ISF402, Iturelix(ORF-23541), JTP-2942, KAI-1455, KAI-1678, KM-9803, KP-101 (GHRP-1),L-346670, L-364343, Labradimil (RMP-7), Lagatide (BN-52080), Lanreotide(ITM-014), Larazotide (AT-1001) (SPD-550), Leconotide (AM-336),Leuprolide (SOT-375), Linaclotide (MD-1100) (MM-41775), Liraglutide(NN-2211), Lixisenatide (AVE-0010) (ZP-10), LSI-518P, Lucinactant,Lusupultide (BY-2001), LY-2189265, LY-2510924, LY-548806, LYN-001,Lypressin, MER-104, Met-enkephalin (INNO-105), Metkephamide (LY-127623),Mifamurtide (CGP-19835) (MLV-19835), MontireIin (CG-3703), MPL-TLB100,MS peptide, MT-li (PT-14), Murabutide (VA-101) (CY-220), Muramyltripeptide, Nafarelin (RS-94991), NBI-6024, Nemifitide (INN-00835),Neogen, Nepadutant (MEN-11420), Nesiritide, Nifalatide (BW942C),NNZ-2566, NP-213, NFC-567, NPY (24-36) (PTL-041120), NT-13, Obinepitide(TM-30338), Octreotide (SMS-201-995), Oglufanide (IM-862), OGP 10-14L,Omiganan (CPI-226), OP-145, ORG-2766 Org-42982 (AG-4263), Ornithinevasopressin, Oxytocin, Ozarelix (D-63153) (SPI-153), p-1025, P-113(PAC-113), Pasireotide (SOM-230), peg-TPOmp (RWJ-800088),Pentigetide(TA-521), Pep-F (5K), Peptide renin inhibitor, Peptide T(AIDS000530), Peptide YY 3-36, Pexiganan (MSI-78), PF-4603629, PI-0824,PI-2301, PL-3994, PLD-116, PMX-53, POL-6326, Posatirelin, PPI-1019,Pralmorelin, Pramlintide, Protirelin, PTH (7-34), PTHrP-(1-36),PTL-0901, PXL-01, R-1516, R-15-K, R-7089, RA peptide, Ramorelix(Hoe-013), RC-3095, Re-188-P-2045 (P2045), rGRF, Romiplostim (AMG-531),Romurtide (DJ-7041), ROSE-010 (GTP-010) (LY-307161), Rotigaptide(ZP-123) (GAP-486), Rusalatide (TP-508), SAN-134, Saralasin (P-113),Secretin (human) (PGN-52) (R-52), Secretin (human) (RG-1068),Semaglutide (NN-9535), SGS-111 Sifuvirtide, SKF-101926, SKF-105494,SKF-110679 (U-75799E), Soblidotin (YHI-501) (TZT-1027), Somatostatin,Somatostatin (D-Trp, D-Cys analog), SP-304 (Guanilib), SPC-3, SPI-1620,SST analog, SUN-11031, SUN-E7001 (CS-872), SYN-1002, Tabilautide(RP-56142), TAK-448, TAK-683, Taltirelin (TA-0910), Tasidotin (ILX-651)(BSF-223651), Taspoglutide (BIM-51077), TCMP-80, Teduglutide(ALX-0600),Teriparatide (LY-333334), Terlakiren (CP-80794), Terlipressin,Tesamorelin (TH-9507), Teverelix (EP-24332), TH-0318, TH-9506,Thymalfasin, Thymodepressin, Thymonoctan (FCE-25388), Thymopentin(TP-5), Thymosin beta-4, Tifuvirtide (R-724) (T-1249), Tigapotide(PCK-3145), Tiplimotide (NBI-5788), TKS-1225 (Oxyntomodulin), TLN-232(CAP-232)(TT-232), TM-30339, TP-9201, TRI-1144, Tridecactide (AP-214),Triletide (Z-420) (ZAMI-420), Triptorelin (WY-42462), TT-223 (El-INT),TT-235, TX14(A), Tyroserleutide (CMS-024), Tyroservatide (CMS-024-02),Ularitide (CDD-95-126) (ESP-305), Unacylated ghrelin (AZP-01) (TH-0332),Urocortin 11, Vapreotide (RC-160), Vasopressin, VIR-576, Xen-2174,XG-102, XOMA-629, Ziconotide (SNX-111), ZP-120, and ZP-1846.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of AC-2592, AC-625,Anaritide, APL-180, Atriopeptin, BGC-728, Carperitide (SUN-4936), CD-NP,CG-77X56, D-4F (APP-018), Danegaptide (ZP-1609) (WAY-261134) (GAP-134),DMP-728 (DU-728), Efegatran (LY-294468), EMD-73495, Eptifibatide(C₆₈-22), ET-642 (RLT-peptide), FE 202158, FX-06, Icatibant (JE-049)(HOE-140), lcrocaptide (ITF-1697), KAI-1455, KM-9803, L-346670,L-364343, LSI-518P, Nesiritide, Peptide renin inhibitor, PL-3994,Rotigaptide (ZP-123) (GAP-486), Saralasin (P-113), SKF-105494,Terlakiren (CP-80794), Tridecactide (AP-214), Ularitide (CDD-95-126)(ESP-305), Urocortin 11, Ziconotide (SNX-111), and ZP-120; and haveutility in the treatment of cardiovascular diseases.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of Azetirelin (YM-14673),Conantokin G, Corticorelin (NEU-3002), CTS-21166 (ASP-1702) (ATG-Z1)(OM-00-3) (OM-99-2), Davunetide (AL-108) (AL-208), Deltibant (CP-0127),Ebiratide (Hoe-427), FGLL, Glypromate, JTP-2942, MontireIin (CG-3703),Nemifitide (INN-00835), NNZ-2566, NT-13, ORG-2766, Peptide T(AIDS000530), Posatirelin, PPI-1019, Protirelin, Secretin (human)(RG-1068), SGS-111, Taltirelin (TA-0910), XG-102, and Ziconotide(SNX-111), and have utility in the treatment of CNS disorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of A-6, Abarelix (PPI-149),ABT-510, ADH-1, AEZS-108 (AN-152) (ZEN-008), Ambamustine (PTT-119),Antagonist G (PTL-68001), ATN-161, Avorelin (EP-23904), Buserelin,Carfilzomib (PR-171), CBP-501, Cemadotin (LU-103793), Chlorotoxin(TM-601), Cilengitide (EMD-121974) (EMD-85189), CTCE-9908, CVX-045,CVX-060, Degarelix (FE 200486), Didemnin B (NSC-325319), DRF-7295,Edotreotide (SMT-487), Elisidepsin (PM-02734), EP-100, Glutoxim(NOV-002), Goralatide (BIM-32001), Goserelin (ICI-118630),Histrelin,Labradimil (RMP-7), Leuprolide (SOT-375), LY-2510924, Met-enkephalin(INNO-105), Mifamurtide (CGP-19835) (MLV-19835), Muramyl tripeptide,Ozarelix (D-63153) (SPI-153), POL-6326, Ramorelix (Hoe-013), RC-3095,Re-188-P-2045 (P2045), Romurtide (DJ-7041), Soblidotin (YHI-501)(TZT-1027), SPI-1620, Tabilautide (RP-56142), TAK-448, TAK-683,Tasidotin (ILX-651) (BSF-223651), Teverelix (EP-24332), Tigapotide(PCK-3145), TLN-232 (CAP-232)(TT-232), Triptorelin (WY-42462),Tyroserleutide (CMS-024), Tyroservatide (CMS-024-02), ZP-1848, inZT0131; and have utility in the treatment of oncological conditions.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of A-623 (AMG-623), AG-284,AI-502, Allotrap 2702 (B-2702), AZD-2315, Cnsnqic-Cyclic (802-2),Delmitide (RDP-58), Dirucotide (MBP-8298) Disitertide (NAFB-001)(P-144), dnaJP1 (AT-001), Edratide (TV-4710), F-991, FAR-404, Glaspimod(SKF-107647), Glatiramer (COP-1), GMDP, IPP-201101, Icatibant (JE049)(HOE-140), MS peptide, Org-42982 (AG-4263), Pentigetide(TA-521),PI-0824, PI-2301, PLD-116, PMX-53, PTL-0901, RA peptide, TCMP-80,Thymodepressin, Thymopentin (TP-5), Tiplimotide (NBI-5788), and ZP-1848;and have utility in the treatment of allergy and immunology disorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of A-71378, AC-162352 (PYY3-36), AC-253, AG2/102, AKL-0707 (LAB GHRH), Albiglutide (GSK-716155),AOD-9604, BAY-73-7977, BIM-44002, BMS-686117, BRX-0585, CJC-1131(DAC:GLP-1), CJC-1134 (PC-DAC) (Exendin-4), CJC-1295 (DAC:GRF),CP-95253, CVX-096 (PF-4856883), Davalintide (AC-2307), Exenatide(AC-2993) (LY-2148568), Exsulin (INGAP Peptide), Glucagon, ISF402,Liraglutide (NN-2211), Lixisenatide (AVE-0010) (ZP-10), LY-2189265,LY-548806, nafarelin (RS 94991), NBI-6024, Obinepitide (TM-30338),Peptide YY 3-36, PF-4603629, Pramlintide, R-7089, Semaglutide (NN-9535),SST analog, SUN-E7001 (CS-872), Taspoglutide (BIM-51077), Tesamorelin(TH-9507), TH-0318, TKS-1225 (Oxyntomodulin), TM-30339, TT-223 (El-INT),Unacylated ghrelin (AZP-01) (TH-0332), and ZT0131, and have utility inthe treatment of metabolic disordrs.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of A-75998, Buserelin,Cetrorelix (NS-75), Detirelix (RS-68439), Ganirelix (Org-37462)(RS-26306), Iturelix, Nafarelin (RS-94991), and triproletin (WY-42462);and have utility in the treatment of fertility.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of AC-100 and p-1025, andhave utility in the treatment of dental disorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of ACV-1, Conantokin G,CJC-1008 (DAC: Dynorphin A), Contulakin G (CGX-1007), CR-665, CR-845,Dynorphin A, E-2078, Felypressin, Frakefamide (LEF-576) (SPD-759)(BCH-3963), HP-228, Icatibant (JE-049) (HOE-140), KAI-1678, Leconotide(AM-336), Metkephamide (LY-127623), MPL-TLB100, NT-13, SYN-1002,TX14(A), Xen-2174, and Ziconotide (SNX-111); and have utility in thetreatment of pain.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of Afamelanotide (EP-1647)(CUV-1647) (Melanotan I), AZX-100, DPK-060, DSC-127, Hemoparatide(PTH(1-37)), Hexapeptide copper II (PC-1358), Pexiganan (MSI-78), PTH(7-34), PXL-01, SKF-110679 (U-75799E), and Thymosin beta-4; and haveutility in the treatment of dermatologic conditions.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of AF-37702, Bivalirudin(BG-8865), carfilomib, (PR-171), CTCE-0214, ETRX 101, H-142, OGP 10-14L,Ornithine vasopressin, peg-TPOmp (RWJ-800088), R-1516, Romiplostim(AMG-531), and TP-9201; and have utility in the treatment of hematologydisorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of Albuvirtide, ALG-889,Alloferon, ALX-40-4C, CB-182804, CB-183315, CZEN-002, Enfuvirtide(T-20), Glucosamyl muramyl tripeptide, Golotimod (SCV-07), GPG-NH2,hLF(1-11), IMX-942, Iseganan (IB-367), Murabutide (VA-101) (CY-220),Neogen, NP-213, Oglufanide (IM-862), Omiganan (CPI-226), OP-145, p-1025,P-113 (PAC-113), Pep-F (5K), R-15-K, Sifuvirtide, SPC-3, Thymalfasin,Thymonoctan (FCE-25388), Tifuvirtide (R-724) (T-1249), TRI-1144,VIR-576, and XOMA-629; and have utility as an antimicrobial or antiviralagent.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of ALTY-0601, B27PD, BDM-E,BIM-23190, CBT-101, Compstatin (POT-4), Eledoisin (ELD-950), andLYN-001, and have utility in the treatment of ophthalmologic disorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of Atosiban (ORF-22164),Barusiban (FE-200400), Carbetocin, Cargutocin (Y-5350), Deslorelin,Oxytocin, and TT-235, and have utility in the treatment of OB-GYNdisorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of Aviptadil (PSD-510),Bremelanotide (PT-141), C-peptide (SPM-933), Desmopressin, EA-230,Lypressin, MER-104, MT-ll (PT-14), SKF-101926, and Vasopressin, and haveutility in the treatment of urologic conditions.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of AC-100, BA-058, Calcitonin(Human), Calcitonin (Salmon), Elcatonin, I-040302 (KUR-112),PTHrP-(1-36), Rusalatide (TP-508), SAN-134, Teriparatide (LY-333334),and ZT031; and have utility in the treatment of bones and connectivetissue disorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of BIO-1211, CGRP (LAB-CGRP),Glucosamyl muramyl tripeptide, GMDP, Icrocaptide (ITF-1697),Lucinactant, Lusupultide (BY-2001), NPC-567, NPY (24-36) (PTL-041120),and Secretin (human) (PGN-52) (R-52); and have utility in the treatmentof respiratory conditions.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of Casokefamide, CCK (25-33),Lagatide (BN-52080), Larazotide (AT-1001) (SPD-550), Linaclotide(MD-1100) (MM-41775), Nepadutant (MEN-11420), Nifalatide (BW942C),ROSE-010 (GTP-010) (LY-307161), Somatostatin, Somatostatin (D-Trp, D-Cysanalog), SP-304 (Guanilib), Teduglutide(ALX-0600), Terlipressin,Triletide (Z-420) (ZAMI-420), Vapreotide (RC-160), ZP-1846, and ZP-1846;and have utility in the treatment of gastroenterologic disorders.

In certain embodiments, compounds of Formula (V) and (VII) are selectedfrom the group consisting of aza-analogues of CJC-1295 (DAC:GRF),DG-3173 (PTR-3173), Dopastatin (BIM-23A760), EP-51216 (EP-51389),Examorelin (EP-23905) (MF-6003), GTP-200 (GTP-300), lpamorelin(NNC-26-0161), Iturelix (ORF-23541), KP-101 (GHRP-1), Lanreotide(ITM-014), Octreotide (SMS-201-995), Pasireotide (SOM-230), Pralmorelin,rGRF, SUN-11031, TH-9506, ZT0131, and vapreotide (RC-160); and haveutility in the treatment of endocrinology disorders.

Compounds of Formula (VI) and (VIII)

Compounds of Formula (VI) and (VIII) are compounds that have utility indrug discovery, diagnosis, treatment and prevention of a disease.

Compounds of Formula (VI) and (VIII) differ from the compounds ofFormula (V), (VII), (IX), and (X) in that compounds of Formula (V),(VII), (IX), and (X) comprise

instead of

at or adjacent to a cleavage and/or a hydrolysis site and/or at theN-terminus and/or the C-terminus of the compound of Formula (VI) or(VIII). R may be selected, e.g., from the group consisting ofunsubstituted and substituted side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, tyrosine, isoleucine (including,R-isoleucine, S-isoleucine and RS-isoleucine), arginine, glycine,asparagine, and glutamine. The side chain radicals may be unsubstitutedor substituted with one or more of the following: a halogen (Cl, F, orBr), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH, —COH, methoxyl,ethoxyl, a C₁-C₆ haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.)or a protecting group (e.g., Phth, Boc, Fmoc, Ddz, etc.).

In certain embodiments, the cleavage site is between the 2^(nd) and3^(rd), 4^(th) and 5^(th), 6^(th) and 7^(th), 7^(th) and 8^(th), 8^(th)and 9^(th) and 10^(th) and 11^(th), 10^(th) and 11^(th), 15^(th) and16^(th), 20^(th) and 21^(st), 24^(th) and 25^(th), 30^(th) and 31^(st),33^(rd) and 34^(th), 36^(th) and 37^(th), 39^(th) and 40^(th), 50^(th)and 51^(st), 24^(th) and 55^(th), 58^(th) and 59^(th), 63^(rd) and64^(th), 66^(th) and 67^(th), 69^(th) and 70^(th), 72^(nd) and 73^(rd),75^(th) and 76^(th), 79^(th) and 80^(th), 82^(nd) and 83^(rd), 85^(th)and 86^(th), 88^(th) and 89^(th), 91^(st) and 99^(th), 105^(th) and106^(th), 114^(th) and 115^(th) amino acid of the peptide (the numberingstarting from the C-terminus of the peptide). In certain embodiments,the cleavage site is Ala-Glu.

Proteins and peptides undergo proteolysis. This process is catalysed bycellular enzymes called proteases. Proteases can be classified intoseven broad groups: (i) serine proteases, (ii) cysteine proteases, (iii)threonine proteases, (iv) aspartic proteases, (v)glutamic proteases,(vi) metalloproteases, and (vii) asparagine peptide lyases. In certainembodiments, the mechanism used to cleave a peptide bond comprisesmaking an amino acid residue that has the cysteine and threonine(proteases) or a water molecule (aspartic acid, metallo- and acidproteases) nucleophilic so that it can attack the peptide carboxylgroup. In certain embodiments, a nucleophile is made by a catalytictriad, where a histidine residue is used to activate serine, cysteine,or threonine as a nucleophile.

Endopeptidase or endoproteinase are proteolytic peptidases that breakpeptide bonds of nonterminal amino acids (i.e. within the molecule), ascompared to exopeptidases, which break peptide bonds from end-pieces ofterminal amino acids. For this reason, endopeptidases cannot break downpeptides into monomers, while exopeptidases can break down proteins intomonomers. A particular case of endopeptidase is the oligopeptidase,whose substrates are oligopeptides instead of proteins. Endopeptidasesare usually very specific for certain amino acids. Endopeptidasesinclude, e.g., trypsin (cuts after Arg or Lys, unless followed by Pro),chymotrypsin (cuts after Phe, Trp, or Tyr, unless followed by Pro; andcuts more slowly after His, Met or Leu), elastase (cuts after Ala, Gly,Ser, or Val, unless followed by Pro), thermolysin (cuts before Ile, Met,Phe, Trp, Tyr, or Val, unless preceded by Pro; and sometimes cuts afterAla, Asp, His or Thr), pepsin (cuts before Leu, Phe, Trp or Tyr, unlesspreceded by Pro; and also others, quite nonspecific), glutamylendopeptidase (cuts after Glu), and neprilysin

An exopeptidase is any peptidase that catalyzes the cleavage of theterminal (or the penultimate) peptide bond. The process releases asingle amino acid or dipeptide from the peptide chain. Depending onwhether the amino acid is released from the amino or the carboxyterminal, an exopeptidase is further classified as an aminopeptidase ora carboxypeptidase, respectively. Thus, an aminopeptidase, an enzyme inthe brush border of the small intestine, will cleave a single amino acidfrom the amino terminal, whereas carboxypeptidase, which is a digestiveenzyme present in pancreatic juice, will cleave a single amino acid fromthe carboxylic end of the peptide.

Cleavage can also take place via i) intra-molecular digestion, ii) lowpH or iii) high temperatures can also cause proteolysisnon-enzymatically.

The cleavage and/or a hydrolysis site of the compound of Formula (VI) orcompound of Formula (VIII) can be determined by one of ordinary skill inthe art without undue experimentation.

In certain embodiments, compounds of Formula (VI) and Formula (VIII)may, e.g., be selected from the group consisting of A-6, blisibimod(A-623), A-71378 (L-Phenylalaninamide,N-(1-oxo-3-(4-(sulfooxy)phenyl)propyl)-L-norleucylglycyl-L-tryptophyl-L-norleucyl-N-methyl-L-alpha-aspartyl),A-75998, Abarelix (PPI-149), ABT-510, AC-100, AC-162352 (PYY 3-36),AC-253, AC-2592, AC-625, ACV-1, ADH-1, AEZS-108 (AN-152) (ZEN-008),AF-37702, Afamelanotide (EP-1647) (CUV-1647) (Melanotan I), AG2/102,AG-284, AI-502, AKL-0707 (LAB GHRH), Albiglutide (GSK-716155),Albuvirtide, ALG-889, Alloferon, Allotrap 2702 (B-2702), ALTY-0601,ALX-40-4C, Ambamustine (PTT-119), Anaritide, Antagonist G (PTL-68001),AOD-9604, APL-180, ATN-161, Atosiban (ORF-22164), Atriopeptin, Aviptadil(PSD-510), Avorelin (EP-23904), AZD-2315, Azetirelin (YM-14673),AZX-100, B27PD, BA-058, Barusiban (FE-200400), BAY-73-7977, BDM-E,BGC-728, BIM-23190, BIM-44002, BIO-1211, Bivalirudin (BG-8865),BMS-686117, Bremelanotide (PT-141), BRX-0585, Buserelin, Calcitonin(Human), Calcitonin (Salmon), Carbetocin, Carfilzomib (PR-171),Cargutocin (Y-5350), Carperitide (SUN-4936), Casokefamide, CB-182804,CB-183315, CBP-501, CBT-101, CCK (25-33), CD-NP, Cemadotin (LU-103793),Cetrorelix (NS-75), CG-77X56, CGRP (LAB-CGRP), Chlorotoxin (TM-601),Cilengitide (EMD-121974) (EMD-85189), CJC-1008 (DAC: Dynorphin A),CJC-1131 (DAC:GLP-1), CJC-1134 (PC-DAC) (Exendin-4), CJC-1295 (DAC:GRF),Cnsnqic-Cyclic (802-2), Compstatin (POT-4), Conantokin G, Contulakin G(CGX-1007), Corticorelin (NEU-3002), CP-95253, C-peptide (SPM-933),CR-665, CR-845, CTCE-0214, CTCE-9908, CTS-21166 (ASP-1702) (ATG-Z1)(OM-00-3) (OM-99-2), CVX-045, CVX-060, CVX-096 (PF-4856883), CZEN-002,D-4F (APP-018), Danegaptide (ZP-1609) (WAY-261134) (GAP-134),Davalintide (AC-2307), Davunetide (AL-108) (AL-208), Degarelix (FE200486), Delmitide (RDP-58), Deltibant (CP-0127), Deslorelin,Desmopressin, Detirelix (RS-68439), DG-3173 (PTR-3173), Didemnin B(NSC-325319), Dirucotide (MBP-8298) Disitertide (NAFB-001) (P-144),DMP-728 (DU-728), dnaJP1 (AT-001), Dopastatin (BIM-23A760), DPK-060,DRF-7295, DSC-127, Dynorphin A, E-2078, EA-230, Ebiratide (Hoe-427),Edotreotide (SMT-487), Edratide (TV-4710), Efegatran (LY-294468),Elcatonin, Eledoisin (ELD-950), Elisidepsin (PM-02734), EMD-73495,Enfuvirtide (T-20), EP-100, EP-51216 (EP-51389), Eptifibatide (C68-22),ET-642 (RLT-peptide), ETRX 101, Examorelin (EP-23905) (MF-6003),Exenatide (AC-2993) (LY-2148568), Exsulin (INGAP Peptide), F-991,FAR-404, FE 202158, Felypressin, FGLL, Frakefamide (LEF-576) (SPD-759)(BCH-3963), FX-06, Ganirelix (Org-37462) (RS-26306), Glaspimod(SKF-107647), Glatiramer (COP-1), Glucagon, Glucosamyl muramyltripeptide, Glutoxim (NOV-002), Glypromate, GMDP, Golotimod (SCV-07),Goralatide (BIM-32001), Goserelin (ICI-118630), GPG-NH2, GTP-200,GTP-300, H-142, Hemoparatide (PTH(1-37)), Hexapeptide copper II(PC-1358), Histrelin, hLF(1-11), HP-228, I-040302 (KUR-112), Icatibant(JE-049) (HOE-140), lcrocaptide (ITF-1697), IMX-942, lpamorelin(NNC-26-0161), IPP-201101, Iseganan (IB-367), ISF402, Iturelix(ORF-23541), JTP-2942, KAI-1455, KAI-1678, KM-9803, KP-101 (GHRP-1),L-346670, L-364343, Labradimil (RMP-7), Lagatide (BN-52080), Lanreotide(ITM-014), Larazotide (AT-1001) (SPD-550), Leconotide (AM-336),Leuprolide (SOT-375), Linaclotide (MD-1100) (MM-41775), Liraglutide(NN-2211), Lixisenatide (AVE-0010) (ZP-10), LSI-518P, Lucinactant,Lusupultide (BY-2001), LY-2189265, LY-2510924, LY-548806, LYN-001,Lypressin, MER-104, Met-enkephalin (INNO-105), Metkephamide (LY-127623),Mifamurtide (CGP-19835) (MLV-19835), MontireIin (CG-3703), MPL-TLB100,MS peptide, MT-li (PT-14), Murabutide (VA-101) (CY-220), Muramyltripeptide, Nafarelin (RS-94991), NBI-6024, Nemifitide (INN-00835),Neogen, Nepadutant (MEN-11420), Nesiritide, Nifalatide (BW942C),NNZ-2566, NP-213, NFC-567, NPY (24-36) (PTL-041120), NT-13, Obinepitide(TM-30338), Octreotide (SMS-201-995), Oglufanide (IM-862), OGP 10-14L,Omiganan (CPI-226), OP-145, ORG-2766 Org-42982 (AG-4263), Ornithinevasopressin, Oxytocin, Ozarelix (D-63153) (SPI-153), p-1025, P-113(PAC-113), Pasireotide (SOM-230), peg-TPOmp (RWJ-800088),Pentigetide(TA-521), Pep-F (5K), Peptide renin inhibitor, Peptide T(AIDS000530), Peptide YY 3-36, Pexiganan (MSI-78), PF-4603629, PI-0824,PI-2301, PL-3994, PLD-116, PMX-53, POL-6326, Posatirelin, PPI-1019,Pralmorelin, Pramlintide, Protirelin, PTH (7-34), PTHrP-(1-36),PTL-0901, PXL-01, R-1516, R-15-K, R-7089, RA peptide, Ramorelix(Hoe-013), RC-3095, Re-188-P-2045 (P2045), rGRF, Romiplostim (AMG-531),Romurtide (DJ-7041), ROSE-010 (GTP-010) (LY-307161), Rotigaptide(ZP-123) (GAP-486)(N-Acetyl-D-tyrosyl-D-prolyl-(4S)-4-hydroxy-D-prolylglycyl-D-alanylglycinamide),Rusalatide (TP-508), SAN-134, Saralasin (P-113), Secretin (human)(PGN-52) (R-52), Secretin (human) (RG-1068), Semaglutide (NN-9535),SGS-111, Sifuvirtide, SKF-101926, SKF-105494, SKF-110679 (U-75799E),Soblidotin (YHI-501) (TZT-1027), Somatostatin, Somatostatin (D-Trp,D-Cys analog), SP-304 (Guanilib), SPC-3, SPI-1620, SST analog,SUN-11031, SUN-E7001 (CS-872), SYN-1002, Tabilautide (RP-56142),TAK-448, TAK-683, Taltirelin (TA-0910), Tasidotin (ILX-651)(BSF-223651), Taspoglutide (BIM-51077), TCMP-80, Teduglutide(ALX-0600),Teriparatide (LY-333334), Terlakiren (CP-80794), Terlipressin,Tesamorelin (TH-9507), Teverelix (EP-24332), TH-0318, TH-9506,Thymalfasin, Thymodepressin, Thymonoctan (FCE-25388), Thymopentin(TP-5), Thymosin beta-4, Tifuvirtide (R-724) (T-1249), Tigapotide(PCK-3145), Tiplimotide (NBI-5788), TKS-1225 (Oxyntomodulin), TLN-232(CAP-232)(TT-232), TM-30339, TP-9201, TRI-1144, Tridecactide (AP-214),Triletide (Z-420) (ZAMI-420), Triptorelin (WY-42462), TT-223 (El-INT),TT-235, TX14(A), Tyroserleutide (CMS-024), Tyroservatide (CMS-024-02),Ularitide (CDD-95-126) (ESP-305), Unacylated ghrelin (AZP-01) (TH-0332),Urocortin 11, Vapreotide (RC-160), Vasopressin, VIR-576, Xen-2174,XG-102, XOMA-629, Ziconotide (SNX-111), ZP-120, ZP-1846, andpharmaceutically acceptable salts thereof.

Synthesis of “Building Blocks” Protected Hydrazide Building BlocksPreparation

Formation of activated hydrazine precursors in solution prior to theirincorporation within a peptide sequence is a part of azapeptidesynthesis. Substituted hydrazines are components in the synthesis, andthe most common protecting groups for substituted hydrazines in solutionand solid phase synthesis of azapeptide are tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl and as well asphthalimide. Two synthetic pathways can be used to prepare the protectedhydrazides: (1) Reduction of protected hydrazones derived from thereaction of carbazate with either aldehyde or ketone; and (2) alkylationof protected hydrazide with an alkyl halide:

wherein R, R₁ and R₂ are as defined above, and PG is protecting group.

Preparation of Phth-Protected Carbamoyl Imidazoles

The compounds of Formula (IA), (TB), (II) and (III) may, e.g., besynthesized by reacting a Phth-protected alkyihydrazine derivative with1,1′-Carbonyldiimidazole (CDI) or 1,1′-carbonyl-bis(3-ethyl imidazolium)triflate (CBET).

Synthesis of the following eight Phth-protected alkyihydrazines wasreported with the following yields:

The following Phth-protected alkylhydrazines can be synthesized by oneof the ordinary skill without undue experimentation, in view of theinformation provided herein and knowledge available in the art:

In certain embodiments, the compounds of Formula (IA), (IB), (II) and(III) may, e.g., be synthesized by reacting a Phth-protectedalkylhydrazine derivative with 1,1′-Carbonyldiimidazole(CDI):

wherein R is selected from the group consisting of side chain radicalsof aspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, and glutamine, and the side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.).

The advantages of this synthetic pathway include, e.g., that Phthalimidegroup bypasses the intramolecular cyclization side product, allows easyaccess to the alkylhydrazine monomers by reduction ofphthaloyl-protected hydrazones derived from N,N-Phthaloylhydrazine witheither an aldehyde or ketone, or, Mitsunobu reaction of alcohol withN-Boc-aminophthalimide, and can be used in both solution and solid phasesyntheses. A 60% hydrazine in DMF for 1-3 hours gave the optimal yieldin the phthaloyl deprotection in both solution and solid phasesyntheses.

The compounds of Formula (IA), (IB), (II) and (III) may also besynthesized by reacting a Phth-protected alkylhydrazine derivative with1,1′-carbonyl-bis(3-ethylimidazolium) triflate (CBEIT) to formcarbamoylimidazolium triflate active building block. CBEIT is anefficient reagent for aminoacylations and peptide couplings:

The following synthetic pathway may be used:

The Phth-protected alkylhydrazine derivative for these syntheses can bemade, e.g., by acidic deprotection of a BOC group ofphthalimide(Phth)-protected N-alkyl-aminophthalimides, thephthalimide(Phth)-protected N-alkyl-aminophthalimide made by Mitsunobureaction of N-tert-butyloxycarbonylaminophthalimide with an appropriatealcohol:

Preparation of Phth-protected Carbamoyl Benzotriazoles

The compounds of Formula (IA), I(B), (II), (III), and (IV) may, e.g., besynthesized by the following scheme:

Wherein R is selected from the group consisting of side chain radicalsof aspartic acid, phenylalanine, alanine, histidine, glutamic acid,tryptophan, valine, leucine, lysine, methionine, tyrosine, isoleucine(including, R-isoleucine, S-isoleucine and RS-isoleucine), arginine,glycine, asparagine, and glutamine, and the side chain radicals may beunsubstituted or substituted with one or more of the following: ahalogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.) or a protecting group (e.g., Phth,Boc, Fmoc, Ddz, etc.).

Synthesis of Azapeptides and Azatides

Compounds of Formula (IA), I(B), (II), (III), and (IV) can be coupled ina linear, stepwise, chain-lengthening fashion to each other, aminoacids, aza-amino acids, peptides, azapeptides, and azatides by eithersolution or liquid phase synthetic methodologies to construct compoundsof Formulas (V) and (VII).

Compounds of Formula (IA), I(B), (II), (III), and (IV) can also be used,e.g., as sub-monomers to elongate and/or cap peptides and azapeptides.

For example, in certain embodiments, compounds of Formula (IA), I(B),(II), (III), and (IV) may be activated by methylation of imidazoleresidue using Mel, and the activated compound may be coupled, e.g., aprotected or unprotected aza-amino acid; a protected or unprotected apeptide; a protected or unprotected azapeptide; a protected orunprotected azatide; or a protected or unprotected compound of Formula(IA), Formula (IB) Formula (II), Formula (III), or Formula (IV); or aprotected or unprotected hydrazine, by either solution or liquid phasesynthetic methodologies, e.g., to form a compound of Formula (V) orFormula (VII). The amino acid, the aza-amino acid, the peptide, theazapeptide, compound of Formula (IA), I(B), (II), (III), and (IV) mayeach be unsubstituted or substituted with one or more of the following:a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g., methyl), hydroxyl, —COOH,—COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆ haloalkyl (e.g., achloromethyl, a fluromethyl, etc.). The methylation of imidazole residuecould, e.g., in acetonitrile at 25° C. under nitrogen for 20 hours.

The methods of the invention may be used to synthesize azapeptides andazatides from 2 to 200 mers in length, e.g., di-azatides, tri-azatides,tetra-azapeptides, penta-azapeptides, etc.

Azapeptide bonds are, e.g., formed by either activation of the hydrazinemoiety or by activation of the N-terminus amine of peptides withcarbonyl donating reagents. After activation, the aza-building blocksare coupled to either a hydrazine moiety or a peptide N-terminus amineto finish the azapeptide bond formation:

Following the bond formation, the peptide elongation is continued byconventional peptide synthesis (either solution or solid phase) untilthe final azapeptide target is reached.

In the methods of the present invention, azapeptides and azatides may beconstructed from hydrazides and peptides with carbonyl donating reagentsinvolving a combination of hydrazine chemistry and peptide synthesis:

wherein R₁ and R₂ is each independently selected from the groupconsisting o side chain radicals of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, and glutamine. Theside chain radicals may be unsubstituted or substituted with one or moreof the following: a halogen (Cl, F, or Br), a C1-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.). In certain embodiments, R₁ andR₂ is each independently selected from the group consisting of H,methyl, isopropyl, isobutyl, benzyl, and side chain radicals of asparticacid, histidine, glutamic acid, tryptophan, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, asparagine, and glutamine.

In certain embodiments, the hydrazide for this synthesis is a compoundof Formula (IA), I(B), (II), (III), or (IV).

In certain embodiments, the carbonyl donor for this synthesis is acompound of Formula (IA), I(B), (II), (III), or (IV).

In certain embodiments, submonomer synthesis of azapeptides comprisesconstructing the azapeptides directly on a solid support using compoundsof Formula (IA), I(B), (II), (III), and (IV). The process comprisesacylation of the solid supported peptides with an activated benzylidenecarbazate, regioselective alkylation of N-terminal semicarbazone, anddeprotection of the semi-carbazone with NH₂OH.HCl, then following theconventional Fmoc-based solid-phase peptide synthesis, elongate theazapeptide to the desired target.

wherein R₁, R₂, R₃, and R₄ is each independently selected from the groupconsisting of side chain radicals of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine (including, R-isoleucine, S-isoleucineand RS-isoleucine), arginine, glycine, asparagine, and glutamine. Theside chain radicals may be unsubstituted or substituted with one or moreof the following: a halogen (Cl, F, or Br), a C₁-C₆ alkyl (e.g.,methyl), hydroxyl, —COOH, —COH, methoxyl, ethoxyl, propoxyl, a C₁-C₆haloalkyl (e.g., a chloromethyl, a fluromethyl, etc.) or a protectinggroup (e.g., Phth, Boc, Fmoc, Ddz, etc.). In certain embodiments, R₁,R₂, R₃, and R₄ is each independently from the group consisting of H,methyl, isopropyl, isobutyl, benzyl, and side chain radicals of asparticacid, histidine, glutamic acid, tryptophan, lysine, methionine,tyrosine, isoleucine (including, R-isoleucine, S-isoleucine andRS-isoleucine), arginine, asparagine, and glutamine.

The submonomer method allows for combinatorial library preparation ofside chain and backbone diverse azapeptides for biological SAR studies.

In certain embodiments, the method of preparing an azapeptide or anazatide comprises hydrolysing a peptide, e.g., a compound of Formula(VI) or (VIII) into fragments and reacting one or more fragments with acompound of Formula (IA), (IB), (II), (III), or (VI).

In certain embodiments, the method of preparing an azapeptide or anazatide comprises cleaving a peptide, e.g., a compound of Formula (VI)or (VIII), into fragments and reacting one or more fragments with acompound of Formula (IA), (IB), (II), (III), or (VI).

In certain embodiments, the method of preparing an azapeptide or anazatide comprises cleaving an end of a peptide, e.g., a compound ofFormula (VI) or (VIII), and reacting the cleaved peptide with a compoundof Formula (IA), (IB), (II), (III), or (VI).

In certain embodiments, the method of preparing an azapeptide or anazatide comprises reacting a compound of Formula (IA), (IB), (II),(III), or (VI) with a truncated peptide.

In certain embodiments, the method of preparing an azapeptide or anazatide comprises conjugating a compound of Formula (IA), (IB), (II),(III), or (VI) with a truncated peptide, e.g., a compound of Formula(VI) or (VII).

In certain embodiments, a method of azapeptide or azatide synthesiscomprises reacting (i) an imidazole derivative of an aza-amino acidcomprising an aza-amino acid covalently bound (conjugated) to aprotecting group at its N-terminus and to imidazole at its C-terminus,wherein the aza-amino acid is selected from the group consisting ofaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, and aza-arginine with (ii) a hydrazide toform an azapeptide. In certain embodiments, the imidazole derivative isa compound of Formula (IA), (IB), (II) and (III).

In certain embodiments, a method of azapeptide or azatide synthesiscomprises reacting (i) an imidazole derivative of an aza-amino acidcomprising an aza-amino acid covalently bound (conjugated) to aprotecting group at its N-terminus and to imidazole at its C-terminus,wherein the aza-amino acid is selected from the group consisting ofaza-glycine, aza-alanine, aza-valine, aza-leucine, aza-isoleucine,aza-proline, aza-phenylalanine, aza-tyrosine, aza-tryptophan,aza-aspartic acid, aza-glutamic acid, aza-aspargine, aza-glutamine,aza-histidine, aza-lysine, and aza-arginine with (ii) a peptide to formthe azapeptide or azatide, wherein the azapeptide or azatide is acompound of formula (V) or (VII).

In certain embodiments, a method of azapeptide or azatide synthesiscompries reacting (i) a benzotriazole derivative of an aza-amino acidcomprising the aza-amino acid covalently bound (conjugated) to aprotecting group at its N-terminus and to benzotriazole at itsC-terminus, wherein the aza-amino acid is selected from the groupconsisting of aza-glycine, aza-alanine, aza-valine, aza-leucine,aza-isoleucine, aza-proline, aza-phenylalanine, aza-tyrosine,aza-tryptophan, aza-aspartic acid, aza-glutamic acid, aza-aspargine,aza-glutamine, aza-histidine, aza-lysine, and aza-arginine with (ii) ahydrazide to form an azapeptide or azatide, wherein the azapeptide orazatide is a compound of formula (V) or (VII).

In certain embodiments, a method of peptide synthesis comprises reacting(i) a benzotriazole derivative of an aza-amino acid comprising theaza-amino acid covalently bound (conjugated) to a protecting group atits N-terminus and to benzotriazole at its C-terminus, wherein theaza-amino acid is selected from the group consisting of aza-glycine,aza-alanine, aza-valine, aza-leucine, aza-isoleucine, aza-proline,aza-phenylalanine, aza-tyrosine, aza-tryptophan, aza-aspartic acid,aza-glutamic acid, aza-aspargine, aza-glutamine, aza-histidine,aza-lysine, and aza-arginine with (ii) a peptide to form an azapeptideor azapeptide, wherein the azapeptide or azatide is a compound offormula (V) or (VII). The protecting group may, e.g., comprisephthalimide, frorenylmethoxycarbonyl, or2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl.

In certain embodiments, the peptide is therapeutically effective for thetreatment of acne, acromegaly, alopecia, anemia, asthma, cancer,age-related macular degeneration, bone cysts, dental caries, cognitiveenhancement, cystic fibrosis, chemoprevention, Cushing's syndrome,anorexia nervosa, depression, obsessive-compulsive disorder, diabeticretinopathy, diabetic macular edema, diabetic nephropathy, dyspepsia,brain edema, epilepsy, renal failure, gingivitis, lupus erythematosus,chronic lyphocytic leukemia, obesity, estrogen deficiency, emesis,endometriosis, endometrial thinning, gastrointestinal disorders,gigantism, bone injuries, tooth restoration, heart failure, myocardialinfarction, cerebrovascular ischemia, ischemia, unstable anginapectoris, hypertension, isolated systolic hypertension, cardiovasculardisease, coronary disorder, atherosclerosis, peripheral artery disease,arrhythmia, pain, vasodilatory hypotension, intradialytic hypotension,stroke, sepsis, thromboembolism, restenosis, hypercalcemia,inflammation, type 1 diabetes, type 2 diabetes, wound healing,eryrthropietic protoporphyria, photodamage, actinic keratosis,myasthenia gravis, multiple sclerosis, transplant rejection, lipidmetabolism disorder, malnutrition, HIV, hepatitis, herpes, glaucoma,osteoporosis, erectile dysfunction, rheumatoid arthritis, Alzheimer'sdisease, dermal scarring, kelid scarring, atopic dermatitis, impetigo,uveitis, uterine contractions, acute coronary syndrome, thrombosis,neutropenia, thrombocytopenia (e.g., heparin-induced thrombocytopenia),female sexual dysfunction, female infertility, postpartum uterineatrophy, postpartum hemorrhage bleeding, Paget's disease, gastricdisorders, Gram negative bacterial infection, mycosesm, bacteremia,candidemia, diarrhea, candida ablicans infection, vulvovaginalcandidiasis, pancreatic dysfunction, benign prostatic hyperplasia,uterine fibroids, growth disorder, metabolic syndrome, metabolicdisorder, HIV-associated lipodystrophy, cachexia, Factor VIIIdeficiency, multiple sclerosis, graft versus host disease, epilepsy,Parkinson's disease, schizophrenia, functional bowel disease,inflammatory bowel disease, irritable bowel syndrome, ulcerativecolitis, Crohn's disease, Celiac disease, short bowel syndrome, ileus,systemic inflammatory response syndrome, brain edema, head injury,precocious puberty, polycystic ovary syndrome, uterine fibroids,nocturia, diabetis insipidus, enuresis, polyuria, primary nocturnalenuresis, Von Willebrand's disease, Hemophilia, hemopoietic disorder,female contraception, male contraception, scleroderma, diabetic footulcer, septic shock, cognition disorder, dementia, HIV-associateddementia, mild cognitive impairment, systemic lupus erythematosus,somatotropin deficiency, muscle wasting, skin disorders, reperfusioninjury, inhibition of premature LH surges, Leukopenia, drug inducedfungal infection, onychomycosis, immune disorder, viral infection,immune deficiency, Huntington's chorea, motor neuron disease,neurodegenerative disorder, psoriasis, tuberculosis, respiratory tractdisorders, postoperative infections, lung disorders, radiation sickness,transplant rejection, hereditary angioedema, rhinitis, allergy, asthma,osteoarthritis, liver cirrhosis, respiratory distress syndrome,stomatitis, pneumonia, nutritional disorders, short stature, respiratorydistress syndrome, lung malformation, postoperative ileus, vasoactiveintestinal peptide, stem cell mobilisation, stem cell transplantation,myelofibrosis, catheter infection, rosacea, otitis, conjunctivitis,neuropathy, control of bleeding, delivery induction, labor initiation,labor stimulation, pemphigus vulgaris, muscle weakness, immunethrombocytopenic purpura, myelodysplastic syndrome, spinal fusion,chronic wounds, bleeding esophageal varices, spinocerebellardegeneration, renal disease, hepatorenal syndrome, insomnia, influenzavirus, aspergillus infection, lung infection, primaryimmunodeficiencies, angiogenesis disorder, recurrent autoimmunecytopenia, decubitus ulcer, varicose ulcer, epidermolysis bullosa, eyesurgery, deafness, or labyrinthitis (inflammation of inner ear).

In certain embodiments, the synthesized azapeptide or azatide istherapeutically effective for the treatment of acne, acromegaly,alopecia, anemia, asthma, cancer, age-related macular degeneration, bonecysts, dental caries, cognitive enhancement, cystic fibrosis,chemoprevention, Cushing's syndrome, anorexia nervosa, depression,obsessive-compulsive disorder, diabetic retinopathy, diabetic macularedema, diabetic nephropathy, dyspepsia, brain edema, epilepsy, renalfailure, gingivitis, lupus erythematosus, chronic lyphocytic leukemia,obesity, estrogen deficiency, emesis, endometriosis, endometrialthinning, gastrointestinal disorders, gigantism, bone injuries, toothrestoration, heart failure, myocardial infarction, cerebrovascularischemia, ischemia, unstable angina pectoris, hypertension, isolatedsystolic hypertension, cardiovascular disease, coronary disorder,atherosclerosis, peripheral artery disease, arrhythmia, pain,vasodilatory hypotension, intradialytic hypotension, stroke, sepsis,thromboembolism, restenosis, hypercalcemia, inflammation, type 1diabetes, type 2 diabetes, wound healing, eryrthropietic protoporphyria,photodamage, actinic keratosis, myasthenia gravis, multiple sclerosis,transplant rejection, lipid metabolism disorder, malnutrition, HIV,hepatitis, herpes, glaucoma, osteoporosis, erectile dysfunction,rheumatoid arthritis, Alzheimer's disease, dermal scarring, kelidscarring, atopic dermatitis, impetigo, uveitis, uterine contractions,acute coronary syndrome, thrombosis, neutropenia, thrombocytopenia(e.g., heparin-induced thrombocytopenia), female sexual dysfunction,female infertility, postpartum uterine atony, postpartum hemorrhagebleeding, Paget's disease, gastric disorders, Gram negative bacterialinfection, mycosesm, bacteremia, candidemia, diarrhea, candida ablicantsinfection, vulvovaginal candidiasis, pancreatic dysfunction, benignprostatic hyperplasia, uterine fibroids, growth disorder, metabolicsyndrome, metabolic disorder, HIV-associated lipodystrophy, cachexia,Factor VIII deficiency, multiple sclerosis, graft versus host disease,epilepsy, Parkinson's disease, schizophrenia, functional bowel disease,inflammatory bowel disease, irritable bowel syndrome, ulcerativecolitis, Crohn's disease, Celiac disease, short bowel syndrome, ileus,systemic inflammatory response syndrome, brain edema, head injury,precocious puberty, polycystic ovary syndrome, uterine fibroids,nocturia, diabetis insipidus, enuresis, polyuria, primary nocturnalenuresis, Von Willebrand's disease, Hemophilia, hemopoietic disorder,female contraception, male contraception, scleroderma, diabetic footulcer, septic shock, cognition disorder, dementia, HIV-associateddementia, mild cognitive impairment, systemic lupus erythematosus,somatotropin deficiency, muscle wasting, skin disorders, reperfusioninjury, inhibition of premature LH surges, Leukopenia, drug inducedfungal infection, onychomycosis, immune disorder, viral infection,immune deficiency, Huntington's chorea, motor neuron disease,neurodegenerative disorder, psoriasis, tuberculosis, respiratory tractdisorders, postoperative infections, lung disorders, radiation sickness,transplant rejection, hereditary angioedema, rhinitis, allergy, asthma,osteoarthritis, liver cirrhosis, respiratory distress syndrome,stomatitis, pneumonia, nutritional disorders, short stature, respiratorydistress syndrome, lung malformation, postoperative ileus, vasoactiveintestinal peptide, stem cell mobilisation, stem cell transplantation,myelofibrosis, catheter infection, rosacea, otitis, conjunctivitis,neuropathy, control of bleeding, delivery induction, labor initiation,labor stimulation, pemphigus vulgaris, muscle weakness, immunethrombocytopenic purpura, myelodysplastic syndrome, spinal fusion,chronic wounds, bleeding esophageal varices, spinocerebellardegeneration, renal disease, hepatorenal syndrome, insomnia, influenzavirus, aspergillus infection, lung infection, primaryimmunodeficiencies, angiogenesis disorder, recurrent autoimmunecytopenia, decubitus ulcer, varicose ulcer, epidermolysis bullosa, eyesurgery, deafness, or labyrinthitis (inflammation of inner ear).

Methods of Treatment

A method of treating acne, acromegaly, alopecia, anemia, asthma, cancer,age-related macular degeneration, bone cysts, dental caries, cognitiveenhancement, cystic fibrosis, chemoprevention, Cushing's syndrome,anorexia nervosa, depression, obsessive-compulsive disorder, diabeticretinopathy, diabetic macular edema, diabetic nephropathy, dyspepsia,brain edema, epilepsy, renal failure, gingivitis, lupus erythematosus,chronic lyphocytic leukemia, obesity, estrogen deficiency, emesis,endometriosis, endometrial thinning, gastrointestinal disorders,gigantism, bone injuries, tooth restoration, heart failure, myocardialinfarction, cerebrovascular ischemia, ischemia, unstable anginapectoris, hypertension, isolated systolic hypertension, cardiovasculardisease, coronary disorder, atherosclerosis, peripheral artery disease,arrhythmia, pain, vasodilatory hypotension, intradialytic hypotension,stroke, sepsis, thromboembolism, restenosis, hypercalcemia,inflammation, type 1 diabetes, type 2 diabetes, wound healing,eryrthropietic protoporphyria, photodamage, actinic keratosis,myasthenia gravis, multiple sclerosis, transplant rejection, lipidmetabolism disorder, malnutrition, HIV, hepatitis, herpes, glaucoma,osteoporosis, erectile dysfunction, rheumatoid arthritis, Alzheimer'sdisease, dermal scarring, kelid scarring, atopic dermatitis, impetigo,uveitis, uterine contractions, acute coronary syndrome, thrombosis,neutropenia, thrombocytopenia (e.g., heparin-induced thrombocytopenia),female sexual dysfunction, female infertility, postpartum uterine atony,postpartum hemorrhage bleeding, Paget's disease, gastric disorders, Gramnegative bacterial infection, mycosesm, bacteremia, candidemia,diarrhea, candida ablicants infection, vulvovaginal candidiasis,pancreatic dysfunction, benign prostatic hyperplasia, uterine fibroids,growth disorder, metabolic syndrome, metabolic disorder, HIV-associatedlipodystrophy, cachexia, Factor VIII deficiency, multiple sclerosis,graft versus host disease, epilepsy, Parkinson's disease, schizophrenia,functional bowel disease, inflammatory bowel disease, irritable bowelsyndrome, ulcerative colitis, Crohn's disease, Celiac disease, shortbowel syndrome, ileus, systemic inflammatory response syndrome, brainedema, head injury, precocious puberty, polycystic ovary syndrome,uterine fibroids, nocturia, diabetis insipidus, enuresis, polyuria,primary nocturnal enuresis, Von Willebrand's disease, Hemophilia,hemopoietic disorder, female contraception, male contraception,scleroderma, diabetic foot ulcer, septic shock, cognition disorder,dementia, HIV-associated dementia, mild cognitive impairment, systemiclupus erythematosus, somatotropin deficiency, muscle wasting, skindisorders, reperfusion injury, inhibition of premature LH surges,Leukopenia, drug induced fungal infection, onychomycosis, immunedisorder, viral infection, immune deficiency, Huntington's chorea, motorneuron disease, neurodegenerative disorder, psoriasis, tuberculosis,respiratory tract disorders, postoperative infections, lung disorders,radiation sickness, transplant rejection, hereditary angioedema,rhinitis, allergy, asthma, osteoarthritis, liver cirrhosis, respiratorydistress syndrome, stomatitis, pneumonia, nutritional disorders, shortstature, respiratory distress syndrome, lung malformation, postoperativeileus, vasoactive intestinal peptide, stem cell mobilisation, stem celltransplantation, myelofibrosis, catheter infection, rosacea, otitis,conjunctivitis, neuropathy, control of bleeding, delivery induction,labor initiation, labor stimulation, pemphigus vulgaris, muscleweakness, immune thrombocytopenic purpura, myelodysplastic syndrome,spinal fusion, chronic wounds, bleeding esophageal varices,spinocerebellar degeneration, renal disease, hepatorenal syndrome,insomnia, influenza virus, aspergillus infection, lung infection,primary immunodeficiencies, angiogenesis disorder, recurrent autoimmunecytopenia, decubitus ulcer, varicose ulcer, epidermolysis bullosa, eyesurgery, deafness, or labyrinthitis (inflammation of inner ear), themethod comprising administering to a subject in need thereof atherapeutically effective amount of a compound of Formula (V) or (VII).

In certain methods, the compound of Formula (V) or (VII) is apeptidomimetic agent comprising a backbone comprising from 2 to 200carbonyl groups and α-nitrogen covalently bound to at least one of saidcarbonyl groups.

In certain methods, the peptidomimetic agent, including compounds ofFormula (V) and (VII), is therapeutically effective for the treatment ofa disorder in the mammal, while a peptide structurally different fromthe compound of Formula (V) and (VII) only in that that the peptidecomprises α-carbon instead of said α-nitrogen is not therapeuticallyeffective.

In certain methods, the compounds of Formula (V) and (VII) have atherapeutic efficacy greater than a peptide structurally different fromthe compounds of Formula (V) and (VII) only in that the peptidecomprises α-carbon instead of said α-nitrogen.

In certain methods, the compounds of Formula (V) and (VII) have longerduration of therapeutic activity than a peptide structurally differentfrom the compounds of Formula (V) and (VII) only in that the peptidecomprises α-carbon instead of said α-nitrogen.

In certain methods, the compounds of Formula (V) and (VII) are moreresistant to protease degradation than a peptide structurally differentfrom the compounds of Formula (V) and (VII) only in that the peptidecomprises α-carbon instead of said α-nitrogen.

In certain methods, the compounds of Formula (V) and (VII) have agreater selectivity to a biological receptor than a peptide structurallydifferent from the compounds of Formula (V) and (VII) only in that thepeptide comprises α-carbon instead of said α-nitrogen.

In certain methods, the compounds of Formula (V) and (VII) have agreater bioavailability than a peptide structurally different from thecompound of Formula (V) and Formula (VII) only in that that the peptidecomprises α-carbon instead of said α-nitrogen.

In certain embodiments, thrombosis is coronary thrombosis.

In certain embodiments, diarrhea is Clostridium difficile-associateddiarrhea.

In certain embodiments, diarrhea is chemotherapy induced diarrhea.

In certain embodiments, respiratory distress syndrome is neonatalrespiratory distress syndrome.

In certain embodiments, respiratory distress syndrome is adult distresssyndrome.

In certain embodiments, said arrhythmia is ventricular arrhythmia.

In certain embodiments, arrhythmia is atrial fibrillation.

In certain embodiments, rhinitis is allergic rhinitis.

In certain embodiments, conjunctivitis is allergic conjunctivitis.

In certain embodiments, osteoporosis is postmenoposal osteoporosis.

In certain embodiments, bone injury is a bone fracture.

In certain embodiments, bacteria is Staphylococcus aureus.

In certain embodiments, neuropathy is diabetic neuropathy.

In certain embodiments, anemia is aplastic anemia, hypoplastic anemia,

In certain embodiments, hepatitis is hepatitis A, hepatitis B orhepatitis C.

Cardiovascular Disorders

A method of treating a cardiovascular disorder comprising administeringa therapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, compounds of Formula (V) are (VII) areaza-analogues of a peptide selected from a group consisting of AC-2592,AC-625, Anaritide, APL-180, Atriopeptin, BGC-728, Carperitide(SUN-4936), CD-NP, CG-77X56, D-4F (APP-018), Danegaptide (ZP-1609)(WAY-261134) (GAP-134), DMP-728 (DU-728), Efegatran (LY-294468),EMD-73495, Eptifibatide (C68-22), ET-642 (RLT-peptide), FE 202158,FX-06, Icatibant (JE-049) (HOE-140), icrocaptide (ITF-1697), KAI-1455,KM-9803, L-346670, L-364343, LSI-518P, Nesiritide, Peptide renininhibitor, PL-3994, Rotigaptide (ZP-123) (GAP-486), Saralasin (P-113),SKF-105494, Terlakiren (CP-80794), Tridecactide (AP-214), Ularitide(CDD-95-126) (ESP-305), Urocortin 11, Ziconotide (SNX-111), and ZP-120.

CNS Disorders

A method of treating a CNS disorder comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodimeents, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting ofAzetirelin (YM-14673), Conantokin G, Corticorelin (NEU-3002), CTS-21166(ASP-1702) (ATG-Z1) (OM-00-3) (OM-99-2), Davunetide (AL-108) (AL-208),Deltibant (CP-0127), Ebiratide (Hoe-427), FGLL, Glypromate, JTP-2942,Montirelin (CG-3703), Nemifitide (INN-00835), NNZ-2566, NT-13, ORG-2766,Peptide T (AIDS000530), Posatirelin, PPI-1019, Protirelin, Secretin(human) (RG-1068), SGS-111, Taltirelin (TA-0910), XG-102, and Ziconotide(SNX-111).

Immune System Disorder

A method of treating an immune system disorder comprising administeringa therapeutically effective amount of a compound of Formula (V) or(VII).

In certain embodiments, the compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting of A-623(AMG-623), AG-284, AI-502, Allotrap 2702 (B-2702), AZD-2315,Cnsnqic-Cyclic (802-2), Delmitide (RDP-58), Dirucotide (MBP-8298)Disitertide (NAFB-001) (P-144), dnaJPl (AT-001), Edratide (TV-4710),F-991, FAR-404, Glaspimod (SKF-107647), Glatiramer (COP-1), GMDP,IPP-201101, Icatibant (JE 049)(HOE-140), MS peptide, Org-42982(AG-4263), Pentigetide(TA-521), PI-0824, PI-2301, PLD-116, PMX-53,PTL-0901, RA peptide, TCMP-80, Thymodepressin, Thymopentin (TP-5),Tiplimotide (NBI-5788), and ZP-1848.

Metabolic Disorders

A method of treating a metabolic disorder comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, the compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting of A-71378,AC-162352 (PYY 3-36), AC-253, AG2/102, AKL-0707 (LAB GHRH), Albiglutide(GSK-716155), AOD-9604, BAY-73-7977, BIM-44002, BMS-686117, BRX-0585,CJC-1131 (DAC:GLP-1), CJC-1134 (PC-DAC) (Exendin-4), CJC-1295 (DAC.GRF),CP-95253, CVX-096 (PF-4856883), Davalintide (AC-2307), Exenatide(AC-2993) (LY-2148568), Exsulin (INGAP Peptide), Glucagon, ISF402,Liraglutide (NN-2211), Lixisenatide (AVE-0010) (ZP-10), LY-2189265,LY-548806, nafarelin (RS 94991), NBI-6024, Obinepitide (TM-30338),Peptide YY 3-36, PF-4603629, Pramlintide, R-7089, Semaglutide (NN-9535),SST analog, SUN-E7001 (CS-872), Taspoglutide (BIM-51077), Tesamorelin(TH-9507), TH-0318, TKS-1225 (Oxyntomodulin), TM-30339, TT-223 (El-INT),Unacylated ghrelin (AZP-01) (TH-0332), and ZT0131.

Fertility

A method of treating fertility comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, the compound of Formula V and (VII) areaza-analogues of a peptide selected from a group consisting of A-75998,Buserelin, Cetrorelix (NS-75), Detirelix (RS-68439), Ganirelix(Org-37462) (RS-26306), Iturelix, Nafarelin (RS-94991), and triproletin(WY-42462).

Dental

A method of treating a dental condition comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, the compound of Formula (V) and (VII) areaza-analogues of AC-100 or p-1025.

Pain

A method of treating pain comprising administering a therapeuticallyeffective amount of a compound of Formula (V) or (VII).

In certain embodiments, the compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting of ACV-1,Conantokin G, CJC-1008 (DAC: Dynorphin A), Contulakin G (CGX-1007),CR-665, CR-845, Dynorphin A, E-2078, Felypressin, Frakefamide (LEF-576)(SPD-759) (BCH-3963), HP-228, Icatibant (JE-049) (HOE-140), KAI-1678,Leconotide (AM-336), Metkephamide (LY-127623), MPL-TLB100, NT-13,SYN-1002, TX14(A), Xen-2174, and Ziconotide (SNX-111).

Dermatology

A method of treating a dermatologic condition comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, the compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting ofafamelanotide (EP-1647) (CUV-1647) (Melanotan I), AZX-100, DPK-060,DSC-127, Hemoparatide (PTH(1-37)), Hexapeptide copper II (PC-1358),Pexiganan (MSI-78), PTH (7-34), PXL-01, SKF-110679 (U-75799E), andThymosin beta-4.

Blood Disorders

A method of treating a blood disorder comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting of AF-37702,Bivalirudin (BG-8865), carfilomib, (PR-171), CTCE-0214, ETRX 101, H-142,OGP 10-14L, Ornithine vasopressin, peg-TPOmp (RWJ-800088), R-1516,Romiplostim (AMG-531), and TP-9201.

Infection

A method of treating an infection comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting ofAlbuvirtide, ALG-889, Alloferon, ALX-40-4C, CB-182804, CB-183315,CZEN-002, Enfuvirtide (T-20), Glucosamyl muramyl tripeptide, Golotimod(SCV-07), GPG-NH2, hLF(1-11), IMX-942, Iseganan (IB-367), Murabutide(VA-101) (CY-220), Neogen, NP-213, Oglufanide (IM-862), Omiganan(CPI-226), OP-145, p-1025, P-113 (PAC-113), Pep-F (5K), R-15-K,Sifuvirtide, SPC-3, Thymalfasin, Thymonoctan (FCE-25388), Tifuvirtide(R-724) (T-1249), TRI-1144, VIR-576, and XOMA-629.

Eye Disorders

A method of treating an eye disorder comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, compounds of Formula (V) or (VII) areaza-analogues of a peptide selected from a group consisting ofALTY-0601, B27PD, BDM-E, BIM-23190, CBT-101, Compstatin (POT-4),Eledoisin (ELD-950), and LYN-001.

Gynecologic Disorder

A method of treating an OB-GYN disorder comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting of Atosiban(ORF-22164), Barusiban (FE-200400), Carbetocin, Cargutocin (Y-5350),Deslorelin, Oxytocin, and TT-235.

Urologic Disorders

A method of treating a urologic disorder comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting of Aviptadil(PSD-510), Bremelanotide (PT-141), C-peptide (SPM-933), Desmopressin,EA-230, Lypressin, MER-104, MT-ll (PT-14), SKF-101926, and Vasopressin.

Bone and Connective Tissue Disorders

A method of treating a bone or a connective tissue disorder comprisingadministering a therapeutically effective amount of a compound ofFormula (V) or (VII) to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) are anaza-analogues of a peptide selected from a group consisting of AC-100,BA-058, Calcitonin (Human), Calcitonin (Salmon), Elcatonin, I-040302(KUR-112), PTHrP-(1-36), Rusalatide (TP-508), SAN-134, Teriparatide(LY-333334), and ZT031.

Respiratory Disorders

A method of treating a respiratory disorder comprising administering atherapeutically effective amount of a compound of Formula (V) or (VII)to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) are anaza-analogues of a peptide selected from a group consisting of BIO-1211,CGRP (LAB-CGRP), Glucosamyl muramyl tripeptide, GMDP, Icrocaptide(ITF-1697), Lucinactant, Lusupultide (BY-2001), NPC-567, NPY (24-36)(PTL-041120), and Secretin (human) (PGN-52) (R-52).

Gastrointestinal Tract Disorders

A method of treating a disorder of a gastrointestinal tract comprisingadministering a therapeutically effective amount of a compound ofFormula (V) or (VII).

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting ofCasokefamide, CCK (25-33), Lagatide (BN-52080), Larazotide (AT-1001)(SPD-550), Linaclotide (MD-1100) (MM-41775), Nepadutant (MEN-11420),Nifalatide (BW942C), ROSE-010 (GTP-010) (LY-307161), Somatostatin,Somatostatin (D-Trp, D-Cys analog), SP-304 (Guanilib),Teduglutide(ALX-0600), Terlipressin, Triletide (Z-420) (ZAMI-420),Vapreotide (RC-160), ZP-1846, and ZP-1846.

Disorders of Endocrine System

A method of treating a disorder of endocrine system comprisingadministering a therapeutically effective amount of a compound ofFormula (V) or (VII) to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from a group consisting of CJC-1295(DAC:GRF), DG-3173 (PTR-3173), Dopastatin (BIM-23A760), EP-51216(EP-51389), Examorelin (EP-23905) (MF-6003), GTP-200 (GTP-300),lpamorelin (NNC-26-0161), Iturelix (ORF-23541), KP-101 (GHRP-1),Lanreotide (ITM-014), Octreotide (SMS-201-995), Pasireotide (SOM-230),Pralmorelin, rGRF, SUN-11031, TH-9506, ZT0131, and vapreotide (RC-160).

Cancer

A method of treating cancer comprising administering a compound ofFormula (V) or (VII) to a subject in need thereof.

In certain embodiments, compounds of Formula (V) and (VII) areaza-analogues of a peptide selected from the group consisting of A-6,Abarelix (PPI-149), ABT-510, ADH-1, AEZS-108 (AN-152) (ZEN-008),Ambamustine (PTT-119), Antagonist G (PTL-68001), ATN-161, Avorelin(EP-23904), Buserelin, Carfilzomib (PR-171), CBP-501, Cemadotin(LU-103793), Chlorotoxin (TM-601), Cilengitide (EMD-121974) (EMD-85189),CTCE-9908, CVX-045, CVX-060, Degarelix (FE 200486), Didemnin B(NSC-325319), DRF-7295, Edotreotide (SMT-487), Elisidepsin (PM-02734),EP-100, Glutoxim (NOV-002), Goralatide (BIM-32001), Goserelin(ICI-118630),Histrelin, Labradimil (RMP-7), Leuprolide (SOT-375),LY-2510924, Met-enkephalin (INNO-105), Mifamurtide (CGP-19835)(MLV-19835), Muramyl tripeptide, Ozarelix (D-63153) (SPI-153), POL-6326,Ramorelix (Hoe-013), RC-3095, Re-188-P-2045 (P2045), Romurtide(DJ-7041), Soblidotin (YHI-501) (TZT-1027), SPI-1620, Tabilautide(RP-56142), TAK-448, TAK-683, Tasidotin (ILX-651) (BSF-223651),Teverelix (EP-24332), Tigapotide (PCK-3145), TLN-232 (CAP-232)(TT-232),Triptorelin (WY-42462), Tyroserleutide (CMS-024), Tyroservatide(CMS-024-02), ZP-1848, and ZT0131.

In certain embodiments, the cancer is breast cancer, colorectal cancer,carcinoid cancers, carcinoma, renal cell carcinoma, endometrialcarcinoma, glioma, glioblastoma, hepatocellular carcinoma, lymphoma,non-small lung cancer, ovarian cancer, gastrointestinal cancer,pancreatic cancer, prostate cancer, sarcoma, solid tumors, metastaticmelanoma, multiple myeloma, malignant melanoma, neuroblastoma, skincancer, non-hodgkin lymphoma, small-cell lung cancer, non-small-lungcancer, mesothelioma, pancreatic cancer, hematological neoplasm,neuroendocrine tumors, pituitary cancer, uterine cancer, orosteosarcoma.

Inflammation

A method of attenuating HMGB1-driven inflammation without impairing theimmune response to microbes with acute lung injury (ALI), comprisingadministering a compound of Formula (V) or (VII) to a subject in needthereof.

Drug Discovery and Development

In certain embodiments, the invention is directed to a method ofincreasing in vivo half-life of a therapeutic peptide, the methodcomprising replacing terminal peptide bonds in the therapeutic peptidewith an azapeptide or azatide linkages, thereby providing a compound ofFormula (V) or (VII).

embodiments, the invention is directed to a method of increasing in vivohalf-life of a therapeutic peptide, the method comprising replacingpeptide bonds adjacent to the terminal peptide bonds in the therapeuticpeptide with an azapeptide or azatide linkages, thereby providing acompound of Formula (V) or (VII).

In certain embodiments, the invention is directed to a method ofincreasing in vivo half-life of a therapeutic peptide, the methodcomprising replacing peptide bonds between the first and second residuesof the therapeutic peptide with an azapeptide or azatide linkages,thereby providing a compound of Formula (V) or (VII).

In certain embodiments, the invention is directed to a method ofimproving therapeutic efficacy of a 2 to 200 amino acid peptide, themethod comprising replacing terminal peptide bonds in the therapeuticpeptide with an azapeptide or azatide linkages, thereby providing acompound of Formula (V) or (VII).

In certain embodiments, the invention is directed to a method ofrendering a peptide therapeutic, the method comprising replacing one ormore amino acids of the peptide with a corresponding aza-amino acid,thereby providing a compound of Formula (V) or (VII).

In additional embodiments, the invention is directed to a drugdevelopment process comprising synthesizing an aza-analogue of a peptidediffering from the peptide in that that an α-carbon of the peptide isreplaced with an α-nitrogen by utilizing compounds of Formula (IA),I(B), (II), (III), and (IV). In some of these embodiments, the analogueis therapeutic, and the peptide is not. In certain embodiments, theaza-analogue has a longer duration of therapeutic activity than thepeptide. In certain embodiments, the aza-analogue analogue has a longerhalf-life than the peptide. In certain embodiments, the aza-analogue hashas an improved efficacy, as compared to the peptide. In certainembodiments, the aza-analogue is more stable to protease degradationthan the peptide. In certain embodiments, the aza-analogue has lessadverse effects than the peptide.

Methods of Improving Efficacy

A method of improving efficacy of a 2 to 50 amino acid peptidecomprising providing an analogue of a peptide, the analogue differingfrom the peptide in that at least one of the amino acids of the peptideis replaced with a corresponding aza-amino acid, wherein the analogue isa compound of Formula (V) or (VII).

Methods of Modulating a Protease Activity

A method of modulating a protease activity comprising exposing thepeptidase to an analogue of a 2 to 50 peptide, the analogue differingfrom the peptide in that at least one of the amino acids of the peptideis replaced with a corresponding aza-amino acid, wherein the analogue isa compound of Formula (V) or (VII).

Methods of Inhibiting a Peptidase

A method of inhibiting a peptidase in a subject comprising administeringan analogue of a peptide to the subject, the analogue differing from thepeptide in that at least one of the amino acids of the peptide isreplaced with a corresponding aza-amino acid, wherein the analogue is acompound of Formula (V) or (VII).

In certain embodiments, the peptidase is an endopeptidase.

In certain embodiments, the peptidase is an exopeptidase.

In certain embodiments, the peptidase is an aspartic protease, aglutamic protease or an asparagine peptide lyase.

In certain embodiments, the peptidase is a retroviral protease.

Pharmaceutical Compositions

Pharmaceutical compositions in accordance with the invention comprise acompound of Formula (V) or Formula (VI) and one or more pharmaceuticallyacceptable excipient(s).

The pharmaceutically acceptable excipients are described in the theHandbook of Pharmaceutical Excipients, Pharmaceutical Press and AmericanPharmacists Association, sixth ed., (2009), incorporated by referenceherein, for all purposes.

The pharmaceutical compositions are designed to be appropriate for theselected mode of administration, and pharmaceutically acceptableexcipients such as, e.g., compatible dispersing agents, buffers,surfactants, preservatives, solubilizing agents, isotonicity agents,stabilizing agents and the like are used as appropriate.

The concentration of the compounds of Formula (V) and Formula (VII) incompositions to be administered is an effective amount and ranges fromas low as about 0.1% by weight to as much as about 95% or about 99.9% byweight.

Typical therapeutically effective amounts or doses can be determined andoptimized using standard clinical techniques and will be dependent onthe mode of administration in view of the information provided hereinand knowledge available in the art.

In certain embodiments, the therapeutically effective amount is a totaldaily dose of from about 0.0003 to about 50 mg of a compound of Formula(V) or (VI) per kg of body weight of the subject.

The pharmaceutical compositions can be formulated, e.g., for oraladministration in solid or liquid form, for parenteral intravenous,subcutaneous, intramuscular, intraperitoneal, intra-arterial, orintradermal injection, for or for vaginal, nasal, topical, or rectaladministration.

Pharmaceutical compositions of the present invention suitable for oraladministration can be presented as discrete dosage forms, e.g., tablets,chewable tablets, caplets, capsules, liquids, and flavored syrups. Suchdosage forms contain predetermined amounts of active ingredients, andmay be prepared by methods of pharmacy well known to those skilled inthe art. See generally, Remington's Pharmaceutical Sciences, 18th ed.,Mack Publishing, Easton Pa. (1990).

Parenteral dosage forms can be administered to subjects by variousroutes including subcutaneous, intravenous (including bolus injection),intramuscular, and intraarterial. Because their administration typicallybypasses patients, natural defenses against contaminants, parenteraldosage forms are specifically sterile or capable of being sterilizedprior to administration to a patient. Examples of parenteral dosageforms include, e.g., solutions ready for injection, dry products readyto be dissolved or suspended in a pharmaceutically acceptable vehiclefor injection, suspensions ready for injection, and emulsions.Pharmaceutical compositions for parenteral injection comprisepharmaceutically acceptable sterile aqueous or nonaqueous solutions,dispersions, suspensions or emulsions and sterile powders forreconstitution into sterile injectable solutions or dispersions.Examples of suitable aqueous and nonaqueous carriers, diluents, solventsor vehicles include water, ethanol, polyols (propylene glycol,polyethylene glycol, glycerol, and the like, and suitable mixturesthereof), vegetable oils (such as olive oil) and injectable organicesters such as ethyl oleate, or suitable mixtures thereof. Suitablefluidity of the composition may be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersions, and by the use of surfactants.These compositions may also contain adjuvants such as preservativeagents, wetting agents, emulsifying agents, and dispersing agents.Prevention of the action of microorganisms may be ensured by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, and the like. It may also bedesirable to include isotonic agents, for example, sugars, sodiumchloride and the like. Prolonged absorption of the injectablepharmaceutical form may be brought about by the use of agents delayingabsorption, for example, aluminum monostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is oftendesirable to slow the absorption of the drug from subcutaneous orintramuscular injection. This may be accomplished by the use of a liquidsuspension of crystalline or amorphous material with poor watersolubility. The rate of absorption of the drug then depends upon itsrate of dissolution which, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of a parenterallyadministered drug form is accomplished by dissolving or suspending thedrug in an oil vehicle.

Suspensions, in addition to the active compounds, may contain suspendingagents, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar, tragacanth, and mixtures thereof.If desired, and for more effective distribution, the compounds of theinvention can be incorporated into slow-release or targeted-deliverysystems such as polymer matrices, liposomes, and microspheres. They maybe sterilized, for example, by filtration through a bacteria-retainingfilter or by incorporation of sterilizing agents in the form of sterilesolid compositions, which may be dissolved in sterile water or someother sterile injectable medium immediately before use.

Injectable depot forms are made by, e.g., forming microencapsulatedmatrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations also are prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues. The injectableformulations can be sterilized, for example, by filtration through abacterial-retaining filter or by incorporating sterilizing agents in theform of sterile solid compositions which can be dissolved or dispersedin sterile water or other sterile injectable medium just prior to use.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic, parenterally acceptablediluent or solvent such as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, one or morecompounds of the invention is mixed, e.g., with at least one inertpharmaceutically acceptable carrier such as sodium citrate or dicalciumphosphate and/or a) fillers or extenders such as starches, lactose,sucrose, glucose, mannitol, and salicylic acid; b) binders such ascarboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia; c) humectants such as glycerol; d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate; e) solutionretarding agents such as paraffin; f) absorption accelerators such asquaternary ammonium compounds; g) wetting agents such as cetyl alcoholand glycerol monostearate; h) absorbents such as kaolin and bentoniteclay; and i) lubricants such as talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof. In the case of capsules, tablets and pills, the dosageform may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using lactose or milk sugar aswell as high molecular weight polyethylene glycols. The solid dosageforms of tablets, dragees, capsules, pills, and granules can be preparedwith coatings and shells such as enteric coatings and other coatingswell known in the pharmaceutical formulating art. They may optionallycontain opacifying agents and can also be of a composition that theyrelease the active ingredient(s) only, or preferentially, in a certainpart of the intestinal tract in a delayed manner. Examples of materialswhich can be useful for delaying release of the active agent can includepolymeric substances and waxes.

Compositions for rectal or vaginal administration include, e.g.,suppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating carriers such as cocoa butter,polyethylene glycol or a suppository wax which are solid at ambienttemperature but liquid at body temperature and therefore melt in therectum or vaginal cavity and release the active compound.

Liquid dosage forms for oral administration include, e.g.,pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents.

Dosage forms for topical or transdermal administration of a compound ofthis invention include, e.g., ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. A desired compound ofthe invention is admixed under sterile conditions with apharmaceutically acceptable carrier and any needed preservatives orbuffers as may be required. The ointments, pastes, creams and gels maycontain, in addition to an active compound of this invention, animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

Powders and sprays can contain, in addition to the compounds of thisinvention, e.g., lactose, talc, silicic acid, aluminum hydroxide,calcium silicates and polyamide powder, or mixtures of these substances.Sprays can additionally contain customary propellants such as, e.g.,chlorofluorohydrocarbons.

Compounds of the invention may also be administered in the form ofliposomes. As is known in the art, liposomes are generally derived fromphospholipids or other lipid substances. Liposomes are formed by mono-or multi-lamellar hydrated liquid crystals that are dispersed in anaqueous medium. Any non-toxic, physiologically acceptable andmetabolizable lipid capable of forming liposomes may be used. Thepresent compositions in liposome form may contain, in addition to thecompounds of the invention, stabilizers, preservatives, and the like.The preferred lipids are the natural and synthetic phospholipids andphosphatidylcholines (lecithins) used separately or together. Methods toform liposomes are known in the art. See, for example, Prescott, Ed.,Methods in Cell Biology, Volume XIV, Academic Press, New York, N.Y.,(1976), p 33 et seq.

Actual dosage levels of active ingredients in the pharmaceuticalcompositions of the invention can be varied so as to obtain an amount ofthe active compound(s) that is effective to achieve the desiredtherapeutic response for a particular subject, compositions and mode ofadministration. The selected dosage level will depend upon the activityof the particular compound, the route of administration, the severity ofthe condition being treated and the condition and prior medical historyof the patient being treated. However, it is within the skill of the artto start doses of the compound at levels lower than required to achievethe desired therapeutic effect and to gradually increase the dosageuntil the desired effect is achieved.

An effective amount of one of the compounds of the invention can beemployed in pure form or, where such forms exist, in pharmaceuticallyacceptable salt form. Alternatively, the compound can be administered asa pharmaceutical composition containing the compound of interest incombination with one or more pharmaceutically acceptable excipient(s).It will be understood, however, that the total daily usage of thecompounds and compositions of the invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient will dependupon a variety of factors including the disorder being treated and theseverity of the disorder; activity of the specific compound employed;the specific composition employed; the age, body weight, general health,sex and diet of the patient; the time of administration, route ofadministration, and rate of excretion of the specific compound employed;the duration of the treatment; the risk/benefit ratio; drugs used incombination or coincidental with the specific compound employed; andlike factors well known in the medical arts. For example, it is wellwithin the skill of the art to start doses of the compound at levelslower than required to achieve the desired therapeutic effect and togradually increase the dosage until the desired effect is achieved.

The total daily dose of the compounds of the present invention asadministered to a human or lower animal may range from about 0.0003 toabout 50 mg/kg of body weight. For purposes of oral administration, morepreferable doses can be in the range of from about 0.0003 to about 5mg/kg body weight. If desired, the effective daily dose can be dividedinto multiple doses for purposes of administration; consequently, singledose compositions may contain such amounts or submultiples thereof tomake up the daily dose. For oral administration, the compositions of theinvention are preferably provided in the form of tablets containingabout 1.0, about 5.0, about 10.0, about 15.0, about 25.0, about 50.0,about 100, about 250, or about 500 milligrams of the compound of Formula(V) or (VII).

Administration

Compounds of Formula (V) and (VII) can be used as therapy to treat avariety of disorders.

Effective doses of the compounds of Formula (V) and Formula (VII), forthe treatment of the above described conditions vary depending upon manydifferent factors, including means of administration, target site,physiological state of the patient, other medications administered, andwhether treatment is diagnostic, prophylactic or therapeutic. One of theordinary skill in the art can determine effective doses of compounds ofFormula (V) and (VII) without undue experimentation.

Administration is performed using standard effective techniques,including, e.g., orally, intravenously, intraperitonealy,subcutaneously, via inhalation, transdermally, intramuscullary,intranasaly, buccaly, sublingualy, or via suppository administration.

Humans amenable to treatment include individuals at risk of disease butnot showing symptoms, as well as patients presently showing symptoms.

Example 1 Preparation of Boc-Protected Alkylhydrazines

Janda's group proposed the preparation of 20 Boc-protectedalkylhydrazines by alkylation of hydrazine with an alkyl halide followedby Boc-protection [56]. Six of the Boc-protected alkylhydrazine monomerswere reported and used as the building blocks to construct diazatides bysolution phase synthesis and Leu-Enkephalin azatides by PEG-supportedliquid phase synthesis:

(Janda, K. D., and Han, H. (1997). Azatide peptidomimetics. (ScrippsResearch Institute, USA; Janda, Kim D.; Han, Hyunsoo.), p. 78).

Example 2 Synthesize of Boc-Protected Hydrazines

Lubell's group synthesized five Boc-protected hydrazines to mimic aminoacid side-chains of Gly, Phe, Val, Ala and Pro:

(Melendez, R. E., and Lubell, W. D. (2004). Aza-Amino Acid Scan forRapid Identification of Secondary Structure Based on the Application ofN-Boc-Azal-Dipeptides in Peptide Synthesis. Journal of the AmericanChemical Society 126, 6759-6764.)

These building blocks were used to synthesize six N-Boc-aza-dipeptidesand then were subsequently introduced into analogues of C-terminalpeptide fragment of human calcitonin gene-related peptide (hCGRP).

Example 3 Preparation of Fmoc-Protected Alkylhydrazines

Lubell's group prepared eleven Fmoc-protected N′-alkylhydrazines bycondensations of Fmoc-protected hydrazine with an appropriate aldehydeor ketone to an acyl hydrazone which was reduced by the catalytichydrogenation and hydride addition:

These monomers were used to construct three biologically active peptidesby partial aza-amino acid scans: the tetrapeptide melanocortin receptor(MCR) agonist, the hexapeptide growth hormone secretagogue (GMIP-6) andthe human calcitonin gene-related peptide (hCGRO) antagonist.

Example 4 Preparation of Ddz-Protected Alkylhydrazines

Gilon's group prepared thirteen Ddz-protected N′-alkylhydrazines bycondensations of commercially available2-(3,5-dimethoxyphenyl)propan-2-yl carbazate (Ddz hydrazine) with eitheraldehyde or ketone to anacyl hydrazone that was reduced by the catalytichydrogenation to yield the desired N′-substituted Ddz hydrazines:

The side chain corresponding to aspartic acid was made by nucleophilicsubstitution of alkyl halide with Ddz hydrazide. Ddz deprotection wasachieved by mild Lewis acid, Mg(ClO4)2, which is orthogonal with the Bocand Fmoc protecting groups and make the solid phase azapeptide synthesiswith Ddz-protected alkylhydrazines compatible with both the Boc and theFmoc strategies.

Freeman, N. S., Hurevich, M., and Gilon, C. (2009). Synthesis ofN′-substituted Ddz-protected hydrazines and their application in solidphase synthesis of aza-peptides. Tetrahedron 65, 1737-1745).

Example 5 Preparation of Phth-Protected Alkylhydrazines

Vanderesse's group prepared seven phthalimide(Phth)-protectedN-alkyl-aminophthalimides by Mitsunobu reaction ofN-tert-butyloxycarbonylaminophthalimide with an appropriate alcohol,followed by the acidic deprotection of the Boc group [59]:

The activated Phth-protected alkylhydrazines monomers were reacted withH-Ala-OMe to give the corresponding aza-dipeptides, which can beintroduced into biologically active peptides and oligomers to formstable p-turn structures:

Example 6 Activation and Coupling of Aza-Amino Acids

Historically, azapeptide bonds are formed by either activation of thehydrazine moiety or by activation of the N-terminus amine of peptideswith carbonyl donating reagents to generate the activated units such asamino isocyanate or isocyanate, activated esters, acid chlorides,carbamoyl imidazole, carbamoyl benzotriazole and1,3,4-oxadiazol-2-(3H)-one, which serve as the activated aza-buildingblocks. The common carbonyl donors used to activate both the hydrazinesand the N-terminus of the growing peptide includingp-nitrophenylchloroformate, bis(2,4-dinitrophenyl) carbonate,bis(pentafluorophenyl) carbonate, N,N′-disuccinimidyl carbonate (DSC),carbonyldiimidazole (CDI), phosgene and triphosgene.

p-Nitrophenylchloroformate

p-Nitrophenylchloroformate was used as the carbonyl donor source toactivate hydrazine. It proved a safer alternative to phosgene andfurnished a more stable activated ester intermediate:

Bis(2,4-dinitrophenyl) carbonate

Bis(2,4-dinitrophenyl) carbonate was used to convert resin-boundN-terminal amino groups into isocyanate, which reacted with a protectedamino acid hydrazide to complete the formation of the azapeptide.Hydantoin formation is the major problem, but can be minimized oreliminated by omission of the base from the activation procedure [45]and/or introducing a reversible amide bond protecting group [61].

Bis(pentafluorophenvl) Carbonate

Bis(pentafluorophenyl) carbonate was used as the carbonyl activatingreagent in preparing azatides [6]. The highly reactivated and easilyhandled carbonyl donor of bis(pentafluorophenyl) carbonate activated awide variety of Boc-protected a-aza-amino acids monomers and coupled tothe azatides. By detailed study, the activated complex was found to bethe amino isocyanate, not the proposed carbamate.

1. Starting from 1-R′-hydrazine Carboxylic Acid,1,1-dimethylethyl Ester

2. Starting from 2-R′-hydrazine Carboxylic Acid,1,1-dimethylethyl Ester

Liley, M., and Johnson, T. (2000). Solid phase synthesis of azapeptidesutilising reversible amide bond protection to prevent hydantoinformation. Tetrahedron Letters 41, 3983-3985.

Other common carbonyl donors failed in coupling reactions due to thecomplicated side reactions, poor reaction yields, and/or prolongedreaction time.

N,N′-disuccinimidyl Carbonate

N,N′-disuccinimidyl carbonate[62, 63] was employed as the carbonylsource to activate benzophenone hydrazone for the synthesis ofaza-glycinyl dipeptides with higher yields and simpler purification:

Carbonyl Diimidazole

Carbonyl diimidazole (CDI) is a useful coupling reagent in amide bondformation, and it also reacted with amines to give stablecarbamoylimidazoles in high yields[64]. Therefore, CDI can be used as acarbonyl donor to activate both amino acid esters and hydrazides inazapeptide formation [65-69]. Azadipeptide synthesis was reported viaactivation of the amino acid ester hydrochloride salts by CDI in thepresence of DIPEA into the active carbamates, which was converted to theisocyanate intermediates. The reactive isocyantes reacted quickly withhydrazide to afford azadipeptides:

(Abo-Dya, N. E., Biswas, S., Basak, A., Avan, I., Alamry, K. A., andKatritzky, A. R. (2013). Benzotriazole-Mediated Synthesis ofAza-peptides: En Route to an Aza-Leuenkephalin Analogue. The Journal ofOrganic Chemistry 78, 3541-3552.)

Solid phase synthesis of Agly peptides was reported by reactingFmoc-NHNH2 with CDI to yield a 1-Fmoc-2-oxoimidazole hydrazinederivative, which reacted with protected peptidyl resin to form theazapeptide resin. The peptide was elongated using standardFmoc/tert-butyl SPPS to the desired target. Final deprotection andcleavage in concentrated TFA furnished Agly peptides with 31 to 53%yield after RP-HPLC purification:

(Mhidia, R., and Melnyk, O. (2010). Selective cleavage of an azaGlypeptide bond by copper(II). Long-range effect of histidine residue.Journal of Peptide Science 16, 141-147)

Phosgene

Commerically available solutions of phosgene in toluene are a convenientsource of carbonyl donors to activate the corresponding L-amino benzylester hydrochloride salts into amino ester isocyanates, which wasinvolved in the aza-amino acid scan with N-Boc-aza-dipeptide strategy.Phosgene solution covered the Fmoc-hydrazine in presence of base to ahighly activated azaglycine building block, the1,3,4-oxadiazol-2(3H)-one. On the other hand, by using anhydrous dioxaneas a solvent and in the absence of a tertiary base, phosgene solutionconverted the Fmoc-methylhydrazines into the corresponding carbazic acidchloride quantitatively at ambient temperature:

The activated Fmoc-aza-amino acid chloride building blocks have beenused to construct Fmoc-Aza-dipeptides in solution phase synthesis:

and the tetrapeptide melanocortin receptor agonist,Ac-His-D-Phe-Arg-Trp-NH2 on solid phase:

Triphosgene

Triphosgene or bis(trichloromethyl) carbonate is a mild, easy-to-handleand efficient carbonylating agent for azapeptide synthesis both insolution and on solid phase. It had been shown to be highly reactive,reducing the reaction temperature and coupling times with high yield andeasy purification [70, 71]. The synthesis of various aza-analogues ofdipeptides, tripeptides and decapeptides has been reported by using bothliquid and solid-phase procedures. (Andre, F., Marraud, M., Tsouloufis,T., Tzartos, S. J., and Boussard, G. (1997). Triphosgene: an efficientcarbonylating agent for liquid and solid-phase aza-peptide synthesis.Application to the synthesis of two aza-analogues of the AChR MIRdecapeptide. Journal of Peptide Science 3, 429-441).

Mel

To a solution of N-phenyl-N-carbamoylimidazole aminophthalimide (0.035mmol) in acetonitrile (0.2 mL) was added Mel (0.175 mmol). The mixturewas stirred at 25° C. under nitrogen for 20 hrs then concentrated todryness and dried under vacuum pump to yield a pale yellow solid(quant.).

Example 7 Submonomer Synthesis of Azapeptides on Solid Phase

The use of activated aza building blocks to generate azapeptides both insolution and on solid phase has been successful for making smallcombinatorial libraries of bio-active compounds for SAR studies. Forexample, this methodology was used for generation of azapeptide analogsof the melanocortin receptor agonist (MCR), the growth hormone releasingpeptide (GHRP-6) and the calcitonin gene-related peptide (CGRP)antagonist for exploring the effect of azapeptide structure onbiological activity [21-25]. However, these are limited in generatingside chain diversity, are difficult to use in preparation of aza aminoacid building blocks, and are subject to contamination with significantside products such as hydantoin and oxadiazalone:

Therefore, a submonomer azapeptide synthesis strategy was introduced toconstruct the aza amino acid residue directly on the SPPS:

The new methodology simplifies azapeptide synthesis and opens the doorfor the combinatorial library preparation of side chain and backbonediverse azapeptides for SAR studies.

Example 8 Submonomer Synthesis of the Growth Hormone Releasing Peptide(GHRP-6) with Side Chain Diversity

Growth hormone releasing peptide 6 (GHRP-6)[His-D-Trp2-Ala3-Trp4-D-Phe5-Lys6-NH2] is a hexapeptide that includesunnatural D-amino acids. GHRP-6 has bio-activity with CD36 receptor andGHS-R1a receptor. Submonomer synthesis produced ten aza-analogs ofGHRP-6 at the D-Trp-Ala-Trp region with yield ranging from 14-42%:

20%

15%

25%

25%

17%

27%

42%

28%

15%

14%

Thirteen aza-arylglycine GHRP-6 analogs were also produced bycopper-catalyzed chemoselective mono-N-arylation of resin-boundsemicarbazone with yields ranging from 0.8-3.4% (FIG. 20):

2.0%

1.0%

1.4%

2.8%

1.4%

1.2%

1.2%

1.0%

1.2%

3.4%

0.8%

1.6%

1.6%Proulx, C., and Lubell, W. D. (2010). Copper-Catalyzed N-Arylation ofSemicarbazones for the Synthesis of Aza-Arylglycine-ContainingAza-Peptides. Organic Letters 12, 2916-2919.

The so-called ‘libraries from libraries’ methodology featured furtherdiversification of the aza-residues and produced seven new GHRP-6azapeptides containing aza-1,2,3-triazole-3-alanine residues by acopper-catalyzed 1,3-dipolar cycloadditiion reaction of aryl azides withaza-proparglycine residues. The isolated yield ranged from 5-11%:

The azapropargyl glycine residues from the submonoer synthesis procedurewere also used for producing the constrained azalysine peptides. Thereaction was accomplished by copper catalyzed coupling of Mannichreagents to azapropargyl glycine residues and eighteen aza-Lys GHRP-6analogs were produced. Zhang, J., Proulx, C., Tomberg, A., and Lubell,W. D. (2014). Multicomponent Diversity-Oriented Synthesis ofAza-Lysine-Peptide Mimics. Organic Letters 16, 298-301.

Example 9 Submonomer Synthesis of Azapeptide Ligands of the InsulinReceptor Tyrosine Kinase (IRTK) with Side Chain Diversity

The pentapeptide, Ac-Asp1-Ile2-Tyr3-Glu4-Thr5-NH2, derived from theactivation loop of IRTK was found to inhibit IRTK phosphorylation. Kato,M., Abe, M., Kuroda, Y., Hirose, M., Nakano, M., and Handa, T. (2009).Synthetic pentapeptides inhibiting autophosphorylation of insulinreceptor in a non-ATP-competitive mechanism. Journal of Peptide Science15, 327-336. 75. Seven azapeptide analogs of the parent pentapeptidewere made by submonomer solid phase synthesis to explore SAR studies onIRTK inhibitory activity in sufficient isolated yield (36-55%).

51%

50%

42%

55%

43%

55%

36%

Example 10 Synthesis of Phthalimide-Protected Carbamoyl Imidazole withPhe-R-Group

To a solution of N-phenyl-aminophthalimide (0.6 mmol) in toluene (2 mL)CDI (1.32 mmol) was added. The mixture was stirred at 90° C. undernitrogen for 20 hours then concentrated to dryness and dried undervacuum pump. The crude product was purified by flash silica gel columnchromatography eluting with hexane/EtOAc (4:3) to afford the products asa white solids 80 mg (38% yield). ¹H NMR (500 MHz, CDCl₃) δ 7.92 (s,1H), 7.79-7.77 (m, 4H), 7.42-7.40 (m, 2H), 7.29-7.28 (m, 3H), 7.22 (dd,1H), 6.95 (d, 1H), 5.03 (s, 2H) ppm. Mass Spectrum: (ESI) m/z 347.27(M+H)⁺.

Example 11 Synthesis of Phthalimide-Protected Carbamoyl Imidazole withZ-Lys-R-Group

To a solution of N-(Z-Lys)-aminophthalimide (0.43 mmol) in toluene (1.5mL) CDI (0.86 mmol) was added. The mixture was stirred at 90° C. undernitrogen for 20 hours then concentrated to dryness and dried undervacuum pump. The crude product was purified by flash silica gel columnchromatography eluting with hexane/EtOAc (4:3) to afford the products asa white solids 60 mg (30% yield). ¹H NMR (500 MHz, CDCl₃) δ 7.81-7.73(m, 5H), 7.28-7.24 (m, 5H), 7.08 (s, 1H), 6.87 (s, 1H), 4.90 (s, 2H),3.78 (t, 2H), 3.16-3.17 (m, 2H), 1.65-1.57 (m, 4H) ppm. Mass Spectrum:(ESI) m/z 347.27 (M+H)⁺.

Example 12 Synthesis of CBEIT

The synthesis of CBEIT was done directly by reaction of CDI with ethyltrifluoromethanesulfonate (triflate) in quantitative yield and no needfor further purification before its use as the coupling reagent.

Example 13

Activation of phthalimide-protected carbamoyl imidazole with Phe-R-groupby Mel

Mel (0.175 mmol) was added to a solution ofN-phenyl-N-carbamoylimidazole aminophthalimide (0.035 mmol) inacetonitrile (0.2 mL). The mixture was stirred at 25° C. under nitrogenfor 20 hours, and, then, concentrated to dryness and dried under vacuumpump to yield a pale yellow solid. The crude product was used directlyfor next step coupling without further purification. Mass Spectrum:(ESI) m/z 361.27 (M*).

Example 14 Coupling Reaction of Activated Phth-Protected CarbamoylImidazole Building Blocks with Protected Amino Acids

O-tert-Butyl-L-serine tert-butyl ester (0.04 mmol) was added to asolution of N-phenyl-N-carbamoylimidazolium iodide aminophthalimide(0.08 mmol) in acetonitrile (0.4 mL). The mixture was stirred at 25° C.under nitrogen for 20 hours and, then, concentrated to dryness and driedunder vacuum pump. The crude product was purified by flash silica gelcolumn chromatography eluting with hexane/EtOAc (4:3) to afford theproducts as a white solid. Mass Spectrum: (ESI) m/z 496.33 (M+H)⁺, m/z518.40 (M+Na)⁺.

L-Phenylalanine tert-butyl ester hydrochloride (0.04 mmol) and DIPEA(0.06 mmol) were added to a solution of N-phenyl-N-carbamoylimidazoliumiodide aminophthalimide (0.08 mmol) in acetonitrile (0.4 mL). Themixture was stirred at 25° C. under nitrogen for 20 hours, and, then,concentrated to dryness and dried under vacuum pump. The crude productwas purified by flash silica gel column chromatography eluting withhexane/EtOAc (4:3) to afford the products as a white solid. MassSpectrum: (ESI) m/z 522.40 (M+H)⁺.

Example 15 Coupling Reaction of Activated Phth-Protected CarbamoylImidazole Building Blocks with Protected Hydrazines

Cbz-hydrazine (0.03 mmol) was added to a solution ofN-phenyl-N-carbamoylimidazolium iodide aminophthalimide (0.03 mmol) inacetonitrile (0.15 mL). The mixture was stirred at 40° C. under nitrogenfor 20 hours then concentrated to dryness and dried under vacuum pump.The crude product was purified by flash silica gel column chromatographyeluting with hexane/EtOAc (4:3) to afford the products as a white solid.Mass Spectrum: (ESI) m/z 445.53 (M+H)⁺, m/z 467.47 (M+Na)⁺.

N-Boc-N-(Z-Lys)-hydrazine (0.03 mmol) was added to a solution ofN-phenyl-N-carbamoylimidazolium iodide aminophthalimide (0.03 mmol) inacetonitrile (0.15 mL). The mixture was stirred at 40° C. under nitrogenfor 20 hours then concentrated to dryness and dried under vacuum pump.The crude product was purified by flash silica gel column chromatographyeluting with hexane/EtOAc (4:3) to afford the products as a white solid.Mass Spectrum: (ESI) m/z 516.53 [(M-Boc)+H]*, m/z 638.33 (M+Na)*.

Example 16

Fmoc-phenylhydrazine was converted into the corresponding carbazic acidchloride by phosgene in anhydrous dichloromethane

and was characterized by X-ray crystallography. X-ray crystallography ofFmoc-phenylhydrazine carbazic acid chloride is depicted in FIG. 13.

Example 17 Synthesis of Di-Azatides by Acid Chloride Coupling

The di-azatides 2 were prepared by coupling of hydrazine 2 with acidchloride in DCM/toluene at 25° C. or 50° C. to yield the N-Fmocprotected di-azatide 1 which was de-protected with piperidine to yieldthe final di-azatide 2:

Another route to the di-azatide 2 was through formation of the protectedcarbazide, and then coupling with an appropriate aldehyde to form anacyl hydrazone which was then reduced by catalytic hydrogenation andhydride addition to di-azatide 3. Chlorosulfonyl isocyanate (CSI) wasthen converted from the amine into the corresponding di-azatide 2:

General Procedure for Coupling of Hydrazine 2 with Acid Chloride (MethodA):

The solution of acid chloride (0.367 mmol) and hydrazine 2 (0.367 mmol)in anhydrous DCM (3 mL) and anhydrous toluene (3 mL) was stirred at 50°C. under nitrogen for 15 hours. The mixture was concentrated to drynessand then the crude product was purified by flash silica gel columnchromatography eluted with hexane/EtOAc mixtures to afford thedi-azatide 1 as white solids or clear oils in 50-70% yield.

General Procedure for Coupling of Protected Carbazide 1 with Aldehydes(Method B):

To a solution of protected carbazide 1 (0.46 mmol) and aldehyde (0.69mmol) in anhydrous methanol (20 mL), triethyl amine (60 uL until pH=7)was added, followed by anhydrous MgSO₄ (200 mg). The mixture was stirredat 55° C. under nitrogen for 1 hour and, then, NaCNBH₃ (2.3 mmol) wasadded, followed by acetic acid (2.3 mmol). The mixture was stirred at80° C. under nitrogen for 15 hours, then concentrated to dryness andpartitioned between water (100 mL) and EtOAc (100 mL). The aqueous layerwas extracted with EtOAc (2×50 mL) and the combined organic phase waswashed with brine (50 mL), dried over Na₂SO₄, filtered and concentratedto afford the crude product which was purified by flash silica gelcolumn chromatography eluted with hexane/EtOAc mixtures to afford thedi-zaatide 3 as white solids or clear oils in 40 to 60% yield.

General Procedure for Coupling of Hydrazine with Acid Chloride:

To a solution of acid chloride (0.367 mmol) and hydrazine (3.67 mmol) inanhydrous DCM (6 mL), N,N,N′,N′-Tetramethyl-1,8-naphthalenediamine(0.734 mmol) was added. The solution was stirred at 25° C. undernitrogen for 15 hours and, then, concentrated to dryness and partitionedbetween 0.5N HCl (20 mL) and EtOAc (20 mL). The aqueous layer wasextracted with EtOAc (2×25 mL) and, then, the combined organic phase waswashed with brine (25 mL), dried over Na₂SO₄, filtered and concentratedto afford the crude product which was purified by flash silica gelcolumn chromatography eluted with hexane/EtOAc mixtures to afford theproducts as white solids or clear oils in 70-80% yield.

General Procedure for Converting Amine into the Corresponding Amide:

To a solution of amine (2 mmol) in anhydrous THE (12 mL) at 0° C.,chlorosulfonyl (CSI) (2 mmol) was added rapidly. The solution wasstirred at 0° C. under nitrogen for 1 hour then water added (10 mL). Thesolution was warmed to room temperature then concentrated andpartitioned between water (100 mL) and EtOAc (100 mL). The aqueous layerwas extracted with EtOAc (2×50 mL) and the combined organic phase waswashed with brine (50 mL), dried over Na₂SO₄, filtered and concentratedto afford the crude product which was purified by flash silica gelcolumn chromatography eluted with hexane/EtOAc mixtures to afford theproducts as white solids in 80-90% yield.

General Procedure for Removing Fmoc Group with Piperidine:

The solution of N-Fmoc protected azatide (1.0 mmol) in piperidine (5 mL)was stirred at 35° C. under nitrogen for 15 mins. The mixture wasconcentrated to dryness and then the crude product was purified by flashsilica gel column chromatography eluted with hexane/EtOAc mixtures toafford the products as white solids in 90-95% yield.

All species were analyzed by ESI mass spectroscopy and selecteddi-azatides were characterized by H and ¹³C NMR:

ESI Mass Spectrum:

                  R =

H Method B (60% yield) m/z 446.13 (M + H⁾⁺ m/z 403.07 (M + H⁾⁺ m/z468.33 (M + Na)⁺ CH₃ Method B (58% yield) m/z 460.13 (M + H⁾⁺ m/z 417.13(M + H⁾⁺ m/z 428.27 (M + Na)⁺ m/z 439.27 (M + Na)⁺

Method A (70% yield) m/z 501.60 (M + H⁾⁺ m/z 523.93 (M + Na)⁺

Method A (55% yield) m/z 520.07 (M + H⁾⁺ m/z 542.20 (M + Na)⁺

Method A (65% yield) m/z 531.60 (M + H⁾⁺ m/z 553.93 (M + Na)⁺

Method B (50% yield) m/z 493.07 (M + H⁾⁺ m/z 515.07 (M + Na)⁺ m/z 535.93(M + H⁾⁺

Method B (45% yield) m/z 532.13 (M + H⁾⁺ m/z 554.33 (M + Na)⁺ m/z 575.07(M + H⁾⁺ m/z 597.20 (M + Na)⁺

Method B (40% yield) m/z 483.27 (M + H⁾⁺ m/z 505.20 (M + Na)⁺

Method B (43% yield) m/z 575.27 (M + H⁾⁺

Method B (42% yield) m/z 509.13 (M + H⁾⁺ m/z 531.20 (M + Na)⁺ Prolineanalogues

Other analogues

Tri-azatide

                R =

H m/z 224.07 (M + H⁾⁺ CH₃ m/z 238.00 (M + H⁾⁺

m/z 280.07 (M + H⁾⁺ m/z 303.20 (M + Na)⁺

m/z 298.00 (M + H⁾⁺ m/z 320.13 (M + Na)⁺

m/z 310.13 (M + H⁾⁺ m/z 332.20 (M + Na)⁺

m/z 314.00 (M + H⁾⁺ m/z 336.00 (M + Na)⁺ Proline analogues

Other analogues

¹H NMR (500 MHz, CD₃OD) δ 7.31-7.23 (m, 7H), 4.99 (br. s, 2H), 4.83 (br.s, 2H), 4.07 (s, 2H) ppm. ¹³CNMR (125 MHz, CD₃OD) δ 159.83, 135.57,130.30, 129.78, 129.39, 54.48 ppm. Mass Spectrum: (ESI) m/z 224.07(M+H)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.49-7.36 (m, 7H), 5.10 (br. s, 2H), 4.95 (br.s, 2H), 4.71 (s, 2H), 3.17 (s, 3H) ppm. ¹³CNMR (125 MHz, CD₃OD) δ163.38, 135.48, 130.24, 129.75, 129.09, 57.49, 39.04 ppm. Mass Spectrum:(ESI) m/z 238.00 (M+H)*.

¹H NMR (500 MHz, CD₃OD) δ 7.40-7.33 (m, 5H), 5.10 (br. s, 2H), 4.69 (s,2H), 4.00-3.00 (br, 2H), 1.90 (m, 1H), 0.94 (d, 6H) ppm. ¹³CNMR (125MHz, CD₃OD) δ 162.90, 137.60, 129.98, 129.44, 129.01, 56.22, 54.51,27.96, 20.64 ppm. Mass Spectrum: (ESI) m/z 280.07 (M+H)⁺, m/z 303.20(M+Na)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.27-7.19 (m, 5H), 4.56 (s, 2H), 3.80-3.60(br., 2H), 3.53 (s, 3H), 2.53 (t, 2H) ppm. ¹³CNMR (125 MHz, CD₃OD) δ174.42, 160.08, 137.61, 129.98, 129.44, 129.01, 54.58, 52.36, 45.75,33.65 ppm. Mass Spectrum: (ESI) m/z 310.13 (M+H)⁺, m/z 332.20 (M+Na)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.38-7.19 (m, 5H), 4.68 (s, 2H), 2.71 (t, 2H)1.98 (s, 3H) ppm. ¹³CNMR (125 MHz, CD₃OD) δ 162.22, 159.98, 137.61,129.89, 129.42, 128.89, 54.50, 48.24, 32.53, 15.18 ppm. Mass Spectrum:(ESI) m/z 298.00 (M+H)⁺, m/z 320.13 (M+Na)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.35-7.28 (m, 10H), 4.63 (br. s, 4H) ppm.¹³CNMR (125 MHz, CD₃OD) δ 164.87, 159.88, 138.43, 138.14, 129.57,129.52, 129.33, 139.23, 128.66, 128.56, 128.46, 128.38, 57.21 ppm. MassSpectrum: (ESI) m/z 314.00 (M+H)⁺, m/z 336.00 (M+Na)⁺.

Example 18

The following Di-azatides were synthesized by following GeneralProcedures described above:

Di-Azatide from N-to-C-Terminal Construction:

Di-Azatide from C-to-N-Terminal Construction:

All species were analyzed by ESI mass spectroscopy and selecteddi-azatides were characterized by ¹H and ¹³C NMR:

¹H NMR (500 MHz, C₃OD) δ 7.31-7.23 (m, 7H),4.99 (br. s, 2H), 4.83 (br.s, 2H), 4.07 (s, 2H) ppm. ¹³C NMR (125 MHz, C₃OD) δ 159.83, 135.57,130.30, 129.78, 129.39, 54.48 ppm. Mass Spectrum: (ESI) m/z 224.07(M+H)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.49-7.36 (m, 7H), 5.10 (br. s, 2H), 4.95 (br.s, 2H), 4.71 (s, 2H), 3.17 (s, 3H) ppm. ¹³CNMR (125 MHz, CD₃OD) δ163.38, 135.48, 130.24, 129.75, 129.09, 57.49, 39.04 ppm. Mass Spectrum:(ESI) m/z 238.00 (M+H)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.40-7.33 (m, 5H), 5.10 (br. s, 2H), 4.69 (s,2H), 4.00-3.00 (br, 2H), 1.90 (m, 1H), 0.94 (d, 6H) ppm. ¹³CNMR (125MHz, CD₃OD) δ 162.90, 137.60, 129.98, 129.44, 129.01, 56.22, 54.51,27.96, 20.64 ppm. Mass Spectrum: (ESI) m/z 280.07 (M+H)⁺, m/z 303.20(M+Na)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.27-7.19 (m, 5H), 4.56 (s, 2H), 3.80-3.60(br., 2H), 3.53 (s, 3H), 2.53 (t, 2H) ppm. ¹³CNMR (125 MHz, CD₃OD) δ174.42, 160.08, 137.61, 129.98, 129.44, 129.01, 54.58, 52.36, 45.75,33.65 ppm. Mass Spectrum: (ESI) m/z 310.13 (M+H)⁺, m/z 332.20 (M+Na)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.38-7.19 (m, 5H), 4.68 (s, 2H), 2.71 (t, 2H)1.98 (s, 3H) ppm. ¹³CNMR (125 MHz, CD₃OD) δ 162.22, 159.98, 137.61,129.89, 129.42, 128.89, 54.50, 48.24, 32.53, 15.18 ppm. Mass Spectrum:(ESI) m/z 298.00 (M+H)⁺, m/z 320.13 (M+Na)⁺.

¹H NMR (500 MHz, CD₃OD) δ 7.35-7.28 (m, 10H), 4.63 (br. s, 4H) ppm.¹³CNMR (125 MHz, CD₃OD) δ 164.87, 159.88, 138.43, 138.14, 129.57,129.52, 129.33, 139.23, 128.66, 128.56, 128.46, 128.38, 57.21 ppm. MassSpectrum: (ESI) m/z 314.00 (M+H)⁺, m/z 336.00 (M+Na)⁺.

Mass Spectrum: (ESI) m/z 684.20 (M+H)⁺, m/z 706.20 (M+Na)⁺.

Example 19

The tri-azatide amide (8) was synthesized by coupling of Di-azatideamide (7) with acid chloride (2) in DCM/toluene at 50° C. to yield theN-Fmoc protected Tri-azatide amide (8), using the General Proceduresdescribed above.

Example 20

Bradykinin amide and four aza-analogues of bradykinin were synthesized:(1)-aza-bradykinin, (8)-aza-bradykin, (1,9)-aza-bradykinin, and(1,8)-aza-bradykin. (1)-aza-bradykinin, (8)-aza-bradykin,(1,9)-aza-bradykinin, and (1,8)-aza-bradykin have the followingstructures:

(1,9)-aza-bradykinin differs from Aza-bradykinin in that the residues atthe N and C terminals have been replaced with corresponding aza-aminoacids.

Aza-Bradykinin (1) Synthesis:

Aza-Bradykinin (1,8) Synthesis:

Aza-Bradykinin (8) Synthesis:

Synthesis of (1,9)-Aza-Bradykinin:

(1,9)-Aza-Bradykinin was synthesized from commercially availablepenta-peptide (Gly-Phe-Ser(OBzl)-Pro-Phe-OtBu) (1) which was coupledwith Fmoc-protected di-proline (2) in present of EDCI, HOBt and DIPEA inDCM to form the hepta-peptide(Fmoc-Pro-Pro-Gly-Phe-Ser(OBzl)-Pro-Phe-OtBu). After the OtBu group washydrolysed in formic acid at 40° C., the acid (3) was further elongatedto octa-azapeptide (5) with Boc-protected Arg(Cbz)₂ hydrazine (4) byapplying TBTU and HOBt coupling reagents. After the Fmoc group wasremoved by NaN₃ in DMF, the free proline (6) was coupled withN-Phth-Arg(Cbz)₂ hydrazine carbazic acid chloride (7) in present ofDIPEA in DCM to form the fully protected aza-bradykinin. N-Boc group wasfirst removed by TFA in DCM, which was then converted into correspondingamide with CSI. Global deprotection of Cbz and OBzl groups was done byapplying catalytic hydrogenation of palladium black and formic acid inmethanol. Finally, the phthaloyl group was removed with a 60% hydrazinein ethanol at 50° C. to yield the (1,9)-aza-bradykinin (K1123W) whichwas purified and isolated by HPLC and characterized by ESI massspectrum.

To make a crude hepta-peptide (3), EDCI (0.04 mmol), HOBt (0.02 mmol)and DIPEA (0.05 mmol) were added to a solution of penta-peptide(Gly-Phe-Ser(OBzl)-Pro-Phe-OtBu) (1) (0.02 mmol) and Fmoc-Pro-Pro-OH (2)(0.023 mmol) in anhydrous DCM (1 mL). The solution was stirred at 25° C.under nitrogen for 2 hours then concentrated to dryness and partitionedbetween water (5 mL) and EtOAc (5 mL). The aqueous layer was extractedwith EtOAc (2×5 mL) and the combined organic phase were washed withbrine (5 mL), dried over Na₂SO₄, filtered and concentrated to afford thecrude product and used directly for next step. Mass Spectrum: (ESI) m/z1116.60 (M+H)⁺, m/z 1138.60 (M+Na)⁺.

The crude hepta-peptide (0.02 mmol) was dissolved in formic acid (1 mL)and the solution was stirred at 35° C. under nitrogen for 3 hours thenconcentrated to dryness and precipitated from MeOH/ether and washed withhexanes to yield white solid. Mass Spectrum: (ESI) m/z 1060.53 (M+H)⁺,m/z 1082.73 (M+Na)⁺.

To a solution of hepta-peptide acid (3) (0.02 mmol) and Boc-protectedArg(Cbz)₂ hydrazine (4) (0.02 mmol) in anhydrous DMF (1 mL) was add TBTU(0.04 mmol), HOBt (0.02 mmol) and DIPEA (0.05 mmol). The solution wasstirred at 40° C. under nitrogen for 2 hrs then concentrated to drynessand wash with water the crude octa-azapeptide was precipitated fromMeOH/ether and washed with hexanes to yield white solid. Mass Spectrum:(ESI) m/z 1542.60 (M+H)⁺, m/z 11564.47 (M+Na)⁺.

To a solution of octa-azapeptide (8.4 umol) in anhydrous DMF (0.3 mL)was added NaN₃ (42 umol). The mixture was stirred at 50° C. undernitrogen for 2 hours then concentrated to dryness and wash with waterthe crude octa-azapeptide amine (6) was precipitated from MeOH/ether andwashed with hexanes to yield white solid. Mass Spectrum: (ESI) m/z1319.67 (M+H)⁺.

To a solution of octa-azapeptide amine (6) (8.4 umol) andN-Phth-Arg(Cbz)₂ hydrazine carbazic acid chloride (7) in anhydrous DCM(0.5 mL) was added DIPEA (2 uL). The mixture was stirred at 25° C. undernitrogen for 15 hrs then concentrated to dryness and wash with water thecrude product was precipitated from MeOH/ether and washed with hexanesto yield white solid. Mass Spectrum: (ESI) m/z 1875.80 (M+H)⁺, m/z1897.40 (M+Na)⁺.

The fully protected aza-bradykinin (6.4 umol) was dissolved in 4N HCl(500 uL). The solution was stirred at 25° C. under nitrogen for 1 hourthen concentrated to dryness and used directly for next step. MassSpectrum: (ESI) m/z 1776.33 (M+H)⁺.

To a solution of the protected aza-bradykinin amine (6.4 umol) inanhydrous THE (0.3 mL) at 0° C. was added chlorosulfonyl (CSI) (9.6umol) rapidly. The solution was stirred at 0° C. under nitrogen for 1hour then water (0.3 mL) was added. The solution was warmed to roomtemperature then concentrated to dryness. The crude product wasprecipitated from MeOH/ether and washed with water to yield white solid.Mass Spectrum: (ESI) m/z 1818.40 (M+H)⁺.

To a solution of the protected aza-bradykinin amide (2.75 umol) inanhydrous MeOH (0.4 mL) was added palladium black (5 mg) and formic acid(100 uL). The mixture was stirred at 60° C. under nitrogen for 5 hoursthen filtered through a pad of celite and concentrated to dryness fornext step. Mass Spectrum: (ESI) m/z 1818.40 (M+H)⁺.

To the crude product in ethanol (500 uL) was added 60% hydrazine (300uL). The solution was stirred at 50° C. for 2 hours and concentrated todryness. The Aza-Bradykinin was purified and isolated by HPLC to yieldwhite solid. Mass Spectrum: (ESI) m/z 1062.67 (M+H)⁺.

Stability of Bradykinin amide and (1-9)-Aza-Bradykinin in serum wasmeasured. The results of stability testing are depicted in Tables 1 and2 and FIG. 1 and FIG. 2.

TABLE 1 Serum: 50 uL; Bradykinin amide: 10 ug O.D. 220 nm (AUC)Incubation time: 1 min, 30 min, 1 hr, 3 hr and 15 hr Bradykinin(10 ug)5305818  1 min 4948859 30 min 2600809  1 hour 2135414  3 hours 155100 15hours 58858

TABLE 2 Serum: 50 uL; Aza-Bradykinin: 5ug O.D. 220 nm (AUC) Incubationtime: 1 min, 30 min, 1 hr, 3 hr and 15 hr Aza-Bradykinin(5 ug) 739361  1min 581369 30 min 578373  1 hour 484321  3 hours 549322 15 hours 521217

As evident from Tables 1 and 2 and FIG. 1 and FIG. 2(1-9)-Aza-Bradykinin is more stable in serum at 1 min, 30 min, 1, hour,3 hours, and 15 hours.

Example 21

Aza-Endomorphin-2 (K763, tetra-azapeptide) of the following structurewas synthesized:

Aza-Endomorphin-2 is an aza-analogue of Endomorphin-2:

Aza-Endomorphin-2 differs from Endomorphin-2 in that the

at the C-terminus and N-terminus of Endormin 2 are replaced with

The following Scheme was used to synthesize Aza-Endomorphin-2:

Commercial available Fmoc-hydrazine (1) was coupled with an4-Methoxybenzaldehyde (2) to form an acyl hydrazone (3) in 90% yield,which was reduced by the catalytic hydrogenation and hydride addition toFmoc protected hydrazide (4) in 90%. The active intermediate acidchloride was generated from carbonyl donor trichloromethyl chloroformate(diphosgene) with the hydrazine (4), which was reacted with excessL-proline to form the azapeptide (5) in 70% (two steps). The key stepwas the coupling reaction between the azapeptide (5) and the diphenylazatide (6) with coupling reagents of TBTU and HOBt in DMF to yield theprotected tetra-azapeptide (7) in 60% yield. Chlorosulfonyl isocyanate(CSI) was then converted the amine into the corresponding amide and theAlBr₃ in EtSH was convert the methoxyl group into hydroxyl amide (8) in67% (two steps). Finally, removed the Fmoc group with piperidine at 35°C. to get the Aza-Endomorphin-2 (K763) in 98% yield.

To a solution of N-Fmoc-4-Methoxyphenylhydrazine (4) (1.06 mmol) andN,N,N′,N′-Tetramethyl-1,8-naphthalenediamine (2.12 mmol) in anhydrousDCM (10 mL) at 0° C. was added dropwise the solution of trichloromethylchloroformate in anhydrous DCM (10 mL). The solution was stirred andwarmed to 25° C. under nitrogen for 30 mins then excess L-prolinesolution in DMF (5.3 mmol) was added. The solution was stirred at 25° C.under nitrogen for 15 hrs then concentrated to dryness and partitionedbetween 0.5N HCl (50 mL) and EtOAc (50 mL). The aqueous layer wasextracted with EtOAc (2×50 mL) and the combined organic phase werewashed with brine (50 mL), dried over Na₂SO₄, filtered and concentratedto afford the crude product which was purified by flash silica gelcolumn chromatography eluting with hexane/EtOAc mixtures to afford theproducts as a white solids in 70% yield. Mass Spectrum: (ESI) m/z 515.93(M+H)⁺, m/z 538.00 (M+Na)⁺.

To a solution of the azapeptide (5) (2.18 mmol) in anhydrous DMF (11 mL)was added TBTU (2.57 mmol) and HOBt (2.18 mmol). The solution wasstirred at room temperature for 10 mins and then added the phenyldiazatide (6) (2.58 mmol) solution in anhydrous DMF (10 mL) at 0° C. wasadded dropwise the solution of trichloromethyl chloroformate inanhydrous DCM (4 mL). The solution was stirred at 25° C. under nitrogenfor 15 hours and, then, concentrated to dryness and partitioned between0.5N HCl (50 mL) and EtOAc (50 mL). The aqueous layer was extracted withEtOAc (2×50 mL) and the combined organic phase were washed with brine(50 mL), dried over Na₂SO₄, filtered and concentrated to afford thecrude product which was purified by flash silica gel columnchromatography eluting with hexane/EtOAc mixtures to afford the productsas a white solids in 60% yield. Mass Spectrum: (ESI) m/z 767.93 (M+H)⁺,m/z 790.07 (M+Na)⁺.

To a solution of the protected tetra-azapeptide (7) (0.52 mmol) inanhydrous THE (12 mL) at 0° C. was added chlorosulfonyl (CSI) (0.78mmol) rapidly. The solution was stirred at 0° C. under nitrogen for 1hour then added water (12 mL). The solution was warmed to roomtemperature then concentrated and partitioned between water (100 mL) andEtOAc (100 mL). The aqueous layer was extracted with EtOAc (2×50 mL) andthe combined organic phase were washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated to afford the crude product which waspurified by flash silica gel column chromatography eluting with 5% MeOHin chloroform to afford the products as a white solids. (ESI) m/z 810.87(M+H)⁺. To a solution of azapeptide amide (0.35 mmol) in EtSH (20 mL) at0° C. was added AlBr₃ solution (1M in DCM) (3.5 mmol). The solution wasstirred and warmed to room temperature under nitrogen for 1.5 hours thenconcentrated to dryness. The crude product which was purified by flashsilica gel column chromatography eluting with acetone: DCM (1:1 v/v) toafford the products as a white solids (67% yield, two steps). (ESI) m/z796.80 (M+H)⁺, m/z 819.00 (M+Na)⁺. The solution of N-Fmoc protectedazapeptide (8) (0.1 mmol) in piperidine (1 mL) was stirred at 35° C.under nitrogen for 15 mins. The mixture was concentrated to dryness andthen the crude product was purified by flash silica gel columnchromatography eluting with acetone: DCM (1:1 v/v) to afford theproducts as a white solids in 98% yield. (ESI) m/z 574.93 (M+H)⁺, m/z597.13 (M+Na)⁺. The aza-Endomorphin (K763) has a similar ¹H and ¹³Cspectra as the Endomorphin-2 (EM2) in pyridine-D₆. Also aza-Endomorphin(K763) has 1:8 cis/trans conformers in DMSO-D₆ by ¹H NMR spectra ofaromatic protons.

Synthesis of Aza-Endomorphin-2 at the proline site (K1167Y)

FIG. 3 depicts ¹H NMR spectra of Endomorphin-2 (EM2) andAza-Endomorphin-2 (K763), respectively (Endomorphin-2 is depicted ontop).

K763 was shown to bind OPRM-1 receptor. The binding is depicted in FIG.10.

A graph of stability of EM2 and K763 in mice serum is depicted in FIG.11.

Pharmacokinetics of K763 at 60 min (IP) with acetonitrile extraction aredepicted in FIG. 12.

FIG. 14A depicts degradation of EM-2, K1167Y and K763 by DPPIV.

FIG. 14B depicts stability of EM-2, K1167Y and K763 in mouse serum.

Example 22

K883, an azapeptide analogue of FSSE, was synthesized. FSSE and K883have the following structures:

K883 differs from FSSE in that the

at the C-terminus and N-terminus of K883 is replaced with

K883 was synthesized in 13 steps; the product of each step was purified,followed by LC-MS to confirm the purity. After purification, compoundswere characterized by high resolution MS and NMR methods (¹H, and ¹³C).

The synthetic procedure was as follows:

Commercially available Fmoc-O-tert-butyl-L-serine (1) and Cbz-hydrazine(2) were coupled in the presence of TBTU, HOBt and DIPEA in DMF to formthe Cbz-protected semicarbazide (3) with 90% yield. The Fmoc group ofthe semicarbazide (3) was removed by sodium azide in DMF to yield thefree amine (4) (86% yield), which was further elongated with anotherFmoc-O-tert-butyl-L-serine (1) in the presence of EDCI, HOBt and DIPEAin DCM to di-Serine Cbz-protected semicarbazide (5) (70% yield). Afterremoval of the Fmoc group with sodium azide in DMF (89% yield), the freeamine (6) was coupled with N-Fmoc-phenyl hydrazine acid chloride (7) inpresent of DIPEA in DCM to form the azapeptide (8) in 90% yield. Then,the Cbz group was de-protected with Pd/C and Et₃SiH in methanol to getthe semicarbazide (9) in 83% yield. Condensations of Fmoc-protectedsemicarbazide (9) with 3-Benzylpropionate aldehyde (10) (Dess Martineoxidation of Benzyl 3-Hydroxypropinonate) to an acyl hydrazone which wasreduced by the catalytic hydrogenation and hydride addition to theprotected aza-tetrapeptide (11) (two steps, 71% yield). CSI thenconverted the amine (11) into the corresponding amide (12) (two steps,60% yield). Fmoc group was removed with sodium azide in DMF to yield thetretra azapeptide (13) (91% yield). De-protection ofO-tert-butyl-L-serine with TFA and O-benzyl with Pd/C and Et₃SiH inmethanol to get the Aza-FSSE (K883) (two steps, 75% yield). There arethirteen reaction steps to the final molecule with the total yield of10%.

To a solution of Cbz-hydrazine (2) (3.0 mmol) andFmoc-O-tert-butyl-L-serine (1) (3.0 mmol) in anhydrous DMF (30 mL) TBTU(3.6 mmol), HOBt (3.0 mmol) and DIPEA (3.0 mmol) were added. Thesolution was stirred at 25° C. under nitrogen for 15 hours, and thenconcentrated to dryness and partitioned between 0.5N HCl (50 mL) andEtOAc (50 mL). The aqueous layer was extracted with EtOAc (2×50 mL) andthe combined organic phase were washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated to afford the crude product which waspurified by flash silica gel column chromatography eluting withhexane/EtOAc mixtures to afford the products as a white solids in 90%yield. ¹H NMR (500 MHz, CDCl₃) δ 8.56 (br, 1H), 7.78 (d, 2H), 7.61 (m,2H), 7.44-7.28 (m, 9H), 6.87 (br, 1H), 5.78 (br, 1H), 5.19 (s, 2H),4.43-4.37 (m, 3H), 4.24 (t, 1H), 3.81 (m, 1H), 3.47 (m, 1H), 1.27 (s,9H) ppm. ¹³CNMR (125 MHz, CDCl₃) δ 170.59, 156.32, 156.12, 144.03,143.81, 141.46, 135.62, 128.73, 128.65, 128.58, 128.37, 127.90, 127.24,125.26, 120.02, 74.85, 68.04, 67.35, 61.42, 53.71, 47.24, 27.53 ppm.Mass Spectrum: (ESI) m/z 532.40 (M+H)⁺, m/z 554.40 (M+Na)⁺.

To a solution of Cbz-protected semicarbazide (3) (5.65 mmol) inanhydrous DMF (30 mL) NaN₃ (6.78 mmol) was added. The mixture wasstirred at 50° C. under nitrogen for 3 hours then concentrated todryness and partitioned between water (50 mL) and EtOAc (50 mL). Theaqueous layer was extracted with EtOAc (2×50 mL) and the combinedorganic phase were washed with brine (50 mL), dried over Na₂SO₄,filtered and concentrated to afford the crude product which was purifiedby flash silica gel column chromatography eluting with hexane/EtOAc/MeOHmixtures to afford the products as a white solids in 86% yield. ¹H NMR(500 MHz, CDCl₃) δ 7.36-7.28 (m, 5H), 6.87 (br, 1H), 5.03 (s, 2H),3.51-3.48 (m, 3H), 1.23 (s, 9H) ppm. ¹³CNMR (125 MHz, CDCl₃) δ 172.83,156.13, 135.80, 128.72, 128.51, 128.35, 73.86, 667.87, 67.35, 63.66,54.87, 27.62 ppm. Mass Spectrum: (ESI) m/z 310.20 (M+H)⁺, m/z 332.20(M+Na)⁺.

To a solution of Cbz-semicarbazide amine (4) (2.9 mmol) andFmoc-O-tert-butyl-L-serine (1) (3.2 mmol) in anhydrous DCM (50 mL) EDCI(4.3 mmol), HOBt (0.58 mmol) and DIPEA (2.9 mmol) were added. Thesolution was stirred at 25° C. under nitrogen for 5 hours thenconcentrated to dryness and partitioned between water (50 mL) and EtOAc(50 mL). The aqueous layer was extracted with EtOAc (2×50 mL) and thecombined organic phase were washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated to afford the crude product which waspurified by flash silica gel column chromatography eluting withhexane/EtOAc mixtures to afford the products as a white solids in 70%yield. ¹H NMR (500 MHz, CDCl₃) δ 8.82 (br, 1H), 7.80 (d, 2H), 7.63 (d,2H), 7.44-7.28 (m, 9H), 7.18 (br, 1H), 6.71 (br, 1H), 5.83 (br, 1H),5.17 (s, 2H), 4.62 (br, 1H) 4.42 (d, 2H), 4.28-4.23 (m, 2H), 3.93 (m,1H), 3.82 (m, 1H), 3.48 (m, 2H), 1.24 (s, 18H) ppm. ¹³CNMR (125 MHz,CDCl₃) δ 170.17, 170.02, 156.21, 155.95, 144.00, 143.81, 141.47, 141.44,135.74, 128.68, 128.50, 128.35, 127.91, 127.25, 125.28, 120.18, 75.30,74.42, 67.90, 67.41, 62.08, 60.80, 60.58, 54.95, 53.03, 47.24, 27.52ppm. Mass Spectrum: (ESI) m/z 675.13 (M+H)⁺, m/z 692.07 (M+Na)⁺.

To a solution of Cbz-protected di-serine semicarbazide (5) (2.49 mmol)in anhydrous DMF (10 mL) was added NaN₃ (3.02 mmol). The mixture wasstirred at 50° C. under nitrogen for 3 hours then concentrated todryness and partitioned between water (25 mL) and EtOAc (25 mL). Theaqueous layer was extracted with EtOAc (2×25 mL) and the combinedorganic phase were washed with brine (25 mL), dried over Na₂SO₄,filtered and concentrated to afford the crude product which was purifiedby flash silica gel column chromatography eluting with hexane/EtOAcmixtures to afford the products as a white solids in 89% yield. MassSpectrum: (ESI) m/z 453.20 (M+H)⁺, m/z 475.33 (M+Na)⁺.

To a solution of Cbz-protected di-serine semicarbazide amine (6) (2.44mmol) and N-Fmoc-phenyl hydrazine acid chloride (7) (2.44 mmol) inanhydrous DCM (24 mL) at 0° C. was added DIPEA (2.44 mmol). The solutionwas stirred and warmed to 25° C. under nitrogen for 5 hours thenconcentrated to dryness and partitioned between water (50 mL) and EtOAc(50 mL). The aqueous layer was extracted with EtOAc (2×50 mL) and thecombined organic phase were washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated to afford the crude product which waspurified by flash silica gel column chromatography eluting withhexane/EtOAc mixtures to afford the products as a white solids in 90%yield. ¹H NMR (500 MHz, CDCl₃) δ 8.93 (br, 1H), 7.80 (d, 2H), 7.54 (d,2H), 7.44 (t, 2H), 7.35-7.31 (m, 12H), 7.18 (br, 2H), 6.63 (br, 1H),6.48 (br, 1H) 6.32 (br, 1H), 5.16 (s, 2H), 4.62 (m, 1H) 4.53 (m, 2H),4.36 (m, 1H), 4.20 (m, 1H), 3.91 (m, 1H), 3.80 (m, 1H), 3.49 (m, 2H),1.20 (s, 18H) ppm. ¹³CNMR (125 MHz, CDCl₃) δ 170.65, 169.92, 157.26,155.90, 155.88, 155.21, 143.27, 143.18, 141.37, 135.62, 135.53, 129.00,128.51, 128.29, 128.14, 128.06, 127.95, 127.20, 124.93, 120.12, 74.94,74.12, 67.70, 61.97, 60.70, 54.76, 52.92, 50.77, 46.96, 27.38 ppm.

To a solution of Cbz-protected di-serine semicarbazide (8) (2.05 mmol)in anhydrous MeOH (10 mL) was added 10% Pd/C (340 mg) followed by Et₃SiH(20.5 mmol). The mixture was stirred at 25° C. under nitrogen for 20mins then filtered through a pad of celite and concentrated to drynessand partitioned between water (50 mL) and EtOAc (50 mL). The aqueouslayer was extracted with EtOAc (2×50 mL) and the combined organic phasewere washed with brine (50 mL), dried over Na₂SO₄, filtered andconcentrated to afford the crude product which was purified by flashsilica gel column chromatography eluting with hexane/EtOAc mixtures toafford the products as a white solids in 83% yield. ¹H NMR (500 MHz,CDCl₃) δ 8.26 (br, 1H), 7.79 (d, 2H), 7.54 (d, 2H), 7.44 (t, 2H),7.35-7.28 (m, 5H), 7.18 (m, 2H), 6.99 (br, 1H), 6.39 (br, 1H), 6.48 (br,1H) 6.30 (d, 1H), 4.50 (m, 3H), 4.37 (m, 1H), 4.20 (m, 1H), 3.91 (m,1H), 3.82 (m, 1H), 3.49-3.39 (m, 2H), 1.26 (s, 9H), 1.18 (s, 9H) ppm.¹³CNMR (125 MHz, CDCl₃) δ 170.73, 170.64, 157.26, 155.34,

143.44, 143.38, 141.59, 135.75, 129.14, 129.08, 128.27, 128.15, 127.39,125.09, 120.31, 75.18, 73.99, 67.95, 62.34, 61.11, 54.80, 53.18, 50.77,47.21, 27.61, 27.58 ppm.

To a solution of protected di-serine semicarbazide (9) (0.87 mmol) and3-Benzylpropionate aldehyde (10) (2.62 mmol) in anhydrous ethanol (6 mL)was added acetic acid (24 uL). The solution was stirred at 25° C. undernitrogen for 2 hrs then concentrated to dryness. The crude productre-dissolved in anhydrous MeOH (20 mL) and added NaCNBH₃ (4.35 mmol)followed by acetic acid (4.35 mmol). The mixture was stirred at 55° C.under nitrogen for 15 hours then concentrated to dryness and partitionedbetween water (50 mL) and EtOAc (50 mL). The aqueous layer was extractedwith EtOAc (2×25 mL) and the combined organic phase were washed withbrine (25 mL), dried over Na₂SO₄, filtered and concentrated to affordthe crude product which was purified by flash silica gel columnchromatography eluting with hexane/EtOAc mixtures to afford the productsas a white solids in 71% yield (two steps). ¹H NMR (500 MHz, CDCl₃) δ8.43 (br, 1H), 7.80 (d, 2H), 7.54 (d, 2H), 7.44 (t, 2H), 7.36-7.31 (m,10H), 7.17 (br, 2H), 7.01 (br, 1H), 6.30 (br, 2H), 5.12 (s, 2H), 4.52(m, 3H), 4.33 (m, 1H), 4.21 (t, 1H), 3.91 (m, 1H), 3.81 (m, 1H), 3.42(m, 2H), 3.14 (t, 2H), 2.56 (t, 2H), 1.22 (s, 9H), 1.17 (s, 9H) ppm. ¹³CNMR (125 MHz, CDCl₃) δ 172.08, 170.59, 169.75, 157.38, 143.40, 143.35,141.56, 136.00, 135.64, 129.13, 129.09, 128.69, 128.41, 128.37, 128.27,128.13, 127.37, 125.08, 125.07, 120.30, 75.10, 73.96, 67.91, 66.54,62.20, 61.11, 60.60, 54.94, 53.23, 50.78, 47.34, 47.15, 32.82, 27.58ppm. Mass Spectrum: (ESI) m/z 851.33 (M+H)⁺, m/z 873.40 (M+Na)⁺.

To a solution of the protected aza-tetrapeptide amine (11) (0.70 mmol)in anhydrous THF (5 mL) at 0° C. was added chlorosulfonyl (CSI) (0.84mmol) rapidly. The solution was stirred at 0° C. under nitrogen for 1hour then added water (5 mL). The solution was warmed to roomtemperature and, then, concentrated and partitioned between water (25mL) and EtOAc (25 mL). The aqueous layer was extracted with EtOAc (2×15mL) and the combined organic phase were washed with brine (25 mL), driedover Na₂SO₄, filtered and concentrated to afford the crude product whichwas purified by flash silica gel column chromatography eluting withhexane/EtOAc mixtures afford the products as a white solids in 60% yield(two steps). ¹H NMR (500 MHz, CDCl₃) δ 9.05 (br, 1H), 7.70 (d, 2H), 7.43(d, 2H), 7.34 (t, 2H), 7.24-7.21 (m, 10H), 7.06 (m, 4H), 6.68 (br, 1H),6.30 (br, 1H), 5.14 (br, 2H), 4.47 (m, 1H), 4.34 9m, 2H), 4.13 (m, 1H),4.09 (m, 2H), 3.82 (m, 2H), 3.76 (m, 2H), 3.63 (m, 2H), 3.51 (m, 1H),3.45 (m, 2H), 3.23 (t, 2H), 1.08 (s, 18H) ppm. ¹³C NMR (125 MHz, CDCl₃)δ 172.10, 170.56, 169.75, 157.36, 157.34, 143.40, 143.35, 141.57,136.03, 136.65, 129.14, 129.09, 128.70, 128.41, 128.37, 128.28, 128.14,127.38, 125.08, 120.31, 75.12, 73.96, 69.94, 66.52, 62.23, 61.13, 54.86,53.24, 50.78, 47.36, 47.10, 32.92, 27.47 ppm. Mass Spectrum: (ESI) m/z894.40 (M+H)⁺, m/z 916.47 (M+Na)⁺.

To a solution of protected aza-tetrapeptide amide (12) (0.48 mmol) inanhydrous DMF (5 mL) was added NaN₃ (0.72 mmol). The mixture was stirredat 50° C. under nitrogen for 2 hours then concentrated to dryness forand partitioned between water (5 mL) and EtOAc (5 mL). The aqueous layerwas extracted with EtOAc (2×5 mL) and the combined organic phase werewashed with brine (5 mL), dried over Na₂SO₄, filtered and concentratedto afford the crude product which was purified by flash silica gelcolumn chromatography eluting with hexane/EtOAc/MeOH mixtures to affordthe products as a white solids in 91% yield. Mass Spectrum: (ESI) m/z672.47 (M+H)⁺, m/z 694.33 (M+Na)⁺.

To a solution of aza-tetrapeptide amide (13) (0.43 mmol) in anhydrousMeOH (5 mL) was added 10% Pd/C (58 mg), followed by Et₃SiH (4.3 mmol).The mixture was stirred at 25° C. under nitrogen for 20 mins thenfiltered through a pad of celite and concentrated to dryness. The acidwas used directly for next step. Mass Spectrum: (ESI) m/z 582.20 (M+H)⁺,m/z 604.27 (M+Na)⁺.

The crude acid (0.43 mmol) was dissolved in l0 mL TFA at 0° C. then wasstirred and warmed to 25° C. under nitrogen for 1 hour. Afterconcentrated to dryness, the crude product was purified by flash silicagel column chromatography eluting with CHC3/MeOH (4:1 v/v) mixture toafford the Aza-FSSE (K883) as a white solids in 75% yield (two steps).¹H NMR (600 MHz, CDCl₃) δ 7.3-7.30 (m, 5H), 4.69 (s, 2H), 4.40 (t, 1H),4.36 (t, 1H), 3.98 (dd, 1H), 3.95 (dd, 1H), 3.85 (dd, 1H), 3.82 (dd,1H), 3.80 (br, 2H), 2.48 (t, 2H) ppm. ¹³CNMR (125 MHz, CDCl₃) δ 178.10,173.33, 170.83, 160.27, 159.81, 136.77, 128.67, 128.07, 127.52, 62.31,61.21, 56.63, 55.42, 52.80, 44.88, 34.91 ppm. Mass Spectrum: (ESI) m/z470.00 (M+H)⁺, m/z 492.07 (M+Na)⁺.

The in vitro and in vivo half-lives of K883 and FSSE were measured. Thein vitro half-life of K883 was greater than 15 hours, while the in vitrohalf-life of the native peptide (FSSE) was 60 min. The in vivo half-lifeof K883 was greater than 69 min, while the in vivo half-life of thenative peptide (FSSE) was less than 1 min. The results are provided inTables 3 and 4 below:

TABLE 3 Individual and Average Plasma Concentrations (ng/ml) for FSSEafter Intravenous Administration at 1 mg/KG in Male Sprague-Dawley ratsIntravenous (1 mg/kg) Rat # Time (hr) 970 971 972 Mean SD 0 (pre-dose)BLOQ BLOQ BLOQ ND ND 0.017 BLOQ BLOQ BLOQ ND ND 0.033 BLOQ BLOQ BLOQ NDND 0.083 BLOQ BLOQ BLOQ ND ND 0.167 BLOQ BLOQ BLOQ ND ND 0.25 BLOQ BLOQBLOQ ND ND 0.33 BLOQ BLOQ BLOQ ND ND 0.50 BLOQ BLOQ BLOQ ND ND AnimalWeight (kg) 0.284 0.271 0.281 0.279 0.007 Volume Dosed (mL) 0.28  0.27 0.28  0.28  0.01  C₀ (ng/mL)¹ ND ND ND ND ND t_(max) (hr)¹ ND ND ND NDND t_(1/2) (hr) ND ND ND ND ND MRT_(last) (hr) ND ND ND ND NO CL(L/hr/kg) ND ND ND ND ND V 

(L/kg) ND ND ND ND ND AUC_(last) (hr · ng/mL) ND ND ND ND ND AUC 

 (hr · ng/mL) ND ND ND ND ND

indicates data missing or illegible when filed

TABLE 4 Individual and Average Plasma Concentrations (ng/ml) for K883after Intravenous Administration at 1 mg/KG in Male Sprague-Dawley RatsIntravenous (1 mg/kg) Rat # Time (hr) 973 974 975 Mean SD 0 (pre-dose)BLOQ BLOQ BLOQ ND ND 0.25 4360    4240    3830 4143 278 0.50 3020   2500    3220 2913 372 1.0  814    1160    1300 1091 250 1.5  281    258    280 273 13.0 2.0  133     130    142 135 6.24 4.0  17.0   15.1 17.1 16.6 1.35 6.0   6.49    4.04  4.77 5.10 1.26 8.0   2.0    0.8711.68 1.52 0.586 Animal   0.288   0.282 0.281 0.284 0.004 Weight (kg)Volume   0.29    0.28  0.28 0.28 0.01 Dosed (mL) C₀ (ng/mL)¹ 6295   7191    4556 6014 1340 t_(max) (hr)¹   0      0    0 0 0 t_(1/2) (hr)  1.30    0.972 1.18 1.15 0.165 MRT_(last) (hr)   0.535   0.528 0.6060.556 0.0430 CL (L/hr/kg)   0.265   0.263 0.267 0.265 0.00200 V 

 (L/kg)   0.144   0.139 0A63 0.149 0.0127 AUC_(last) 3772    3807   3749 3776 29.4 (hr · ng/mL) AUC 

  3776    3808    3751 3778 28.5 (hr · ng/mL)

indicates data missing or illegible when filed

Both K883 and FSSE were tested in animal studies and shown to beprotective.

The results of stability and pK study of FSSE and K833 in mice aredepicted in FIGS. 4 and 5.

The results of Ac-FSSE and K883 binding to MD-2 study are depicted inFIG. 6.

The results of Ac-FSSE and K883 inhibiting MD-2 binding to HMGB1 aredepicted in FIG. 6.

FIG. 7 depicts graphs showings that K883 inhibits HMGB1-induced TNFsecretion.

Example 23 Synthesis of Phth-Phe-Aa-Di-Azapeptide

Phth-Phe-Aa-di-azapeptide was synthesized using the following reactionand conditions:

The yield was 70%.

Example 24 Synthesis of Phth-Phe-Aa-di-azapeptide

Phth-Phe-Aa-di-azapeptide was synthesized using the following reactionand conditions:

The yield was 80%.

In the preceding specification, the invention has been described withreference to specific exemplary embodiments and examples thereof. Itwill, however, be evident that various modifications and changes may bemade thereto without departing from the broader spirit and scope of theinvention as set forth in the claims that follow. The specification anddrawings are accordingly to be regarded in an illustrative manner ratherthan a restrictive sense. All documents cited herein, as well as textappearing in the figures, are hereby incorporated by reference in theirentirety for all purposes to the same extent as if each were soindividually denoted.

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What is claimed is:
 1. A compound of Formula (IA):

wherein X is imidazolyl or benzotriazolyl, and wherein (i) A and R areconnected and form a side chain of proline, or (ii) A is hydrogen, or aprotecting group comprising phthalimidyl, tert-butoxycarbonyl,9-fluorenylmethoxycarbonyl,2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl; and R is selected fromthe group consisting of side chain radicals of aspartic acid,phenylalanine, alanine, histidine, glutamic acid, tryptophan, valine,leucine, lysine, methionine, tyrosine, isoleucine, arginine, glycine,asparagine, serine, cysteine, serine, threonine, and glutamine.
 2. Thecompound of claim 1, wherein R is substituted with one or more of thefollowing: a halogen, a C₁-C₆ alkyl, hydroxyl, —COOH, —COH, methoxyl,ethoxyl, propoxyl, a C₁-C₆ haloalkyl or a protecting group.
 3. Thecompound of claim 1, wherein R is unsubstituted.
 4. The compound ofclaim 1, wherein X is imidazolyl.
 5. The compound of claim 4, which isselected from the group consisting of

wherein PG is selected from the group consisting of H, phthalimidyl,tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, and2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl.
 6. The compound of claim1, wherein X is benzotriazolyl.
 7. The compound of claim 6, wherein thecompound is selected from the group consisting of

wherein PG is selected from the group consisting of H, phthalimidyl,tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, and2-(3,5-dimethoxyphenyl)propan-2-yloxycarbonyl.
 8. A method of synthesisof an azapeptide comprising coupling a compound according to claim 1 toan amino acid or an aza-amino acid, wherein the azapeptide is a compoundof Formula (V):

or a pharmaceutically acceptable salt thereof, wherein B isindependently selected from the group consisting of hydrogen, —NH₂,—NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄ alkyl, —COC₁-C₄ haloalkyl, —OH, anamino acid, an aza amino acid, a 2 to 60-mer peptide, a 2 to 60-mer azapeptide, a 2 to 60-mer azatide; D is independently selected from thegroup consisting of —OH, —NH₂, —NNH₂, —CONH₂, —COOH, —COH, —COC₁-C₄alkyl, —COC₁-C₄ haloalkyl, an amino acid, an aza amino acid, a 2 to60-mer peptide, a 2 to 60-mer aza peptide, a 2 to 60-mer azatide; and Ris independently selected from the group consisting of unsubstituted andsubstituted side chain radicals of aspartic acid, phenylalanine,alanine, histidine, glutamic acid, tryptophan, valine, leucine, lysine,methionine, tyrosine, isoleucine, arginine, glycine, asparagine, serine,and glutamine.
 9. The method of claim 8, wherein the coupling is duringa solid phase peptide synthesis.
 10. The method of claim 8, wherein thecoupling is during a liquid phase peptide synthesis.
 11. The method ofclaim 9, comprising activating the compound according to claim 1 priorto the coupling.
 12. The method of claim 11, wherein the activating iswith Mel.
 13. The method of claim 11, wherein the activating is withDIPEA.
 14. The method of claim 10, comprising activating the compoundaccording to claim 1 prior to the coupling.
 15. The method of claim 14,wherein the activating is with Mel.
 16. The method of claim 14, whereinthe activating is with DIPEA.
 17. The method of claim 8, wherein thecompound of Formula (V) is a di-azatide, a tri-azatide, atetra-azapeptide, or a tetra-azatide.
 18. The method of claim 9, whereinthe compound of Formula (V) is produced in a yield of at least about50%.
 19. The method of claim 10, wherein the compound of Formula (V) isproduced in a yield of at least about 50%.
 20. A method of treating adisorder comprising administering a therapeutically effective amount ofthe azapeptide prepared according to claim 9 or claim 10 to a subject inneed thereof.